A breeding method for enoki mushrooms

Abstract

This invention relates to the field of edible fungi technology, providing a method for breeding enoki mushrooms. The method includes: collecting enoki mushroom strain samples from different regions; conducting morphological observations on the collected strains; culturing the strains on standard culture media; testing the strains' resistance to diseases and pests; selecting strains with fast growth rates, high bioconversion rates, and strong resistance to diseases and pests for hybridization; screening the hybrid offspring to select individuals with round caps, straight stems, and pure white color; purifying the selected individuals through self-pollination; and expanding the cultivation of the finally selected superior strains for wider application and promotion. Through precise control of environmental parameters and strict selection criteria, enoki mushroom varieties with superior traits can be bred more efficiently and accurately, resulting in significant improvements in growth rate, morphological quality, and stress resistance.

Description

Technical Field

[0001] This invention relates to the field of edible fungi technology, specifically to a method for breeding enoki mushrooms. Background Technology

[0002] Enoki mushrooms, a popular edible fungus, hold an important position in the global agricultural industry. However, traditional cultivated varieties often face problems such as low yields and high susceptibility to pests and diseases, which seriously affect the yield and quality of enoki mushrooms and limit the further development of the industry.

[0003] In the cultivation and production of enoki mushrooms, improving quality and yield are key factors in enhancing market competitiveness. Traditional enoki mushroom breeding techniques mainly rely on natural selection and artificial hybridization. These methods are not only time-consuming but also inefficient, making it difficult to meet the rapidly evolving market demands. With advancements in biotechnology, it has become possible to improve varieties using modern biotechnologies such as molecular marker-assisted selection and genetic engineering. However, these technologies often require high technical barriers and costs, limiting their widespread application in some regions.

[0004] Therefore, developing a breeding method for enoki mushrooms that combines traditional and modern technologies can utilize the stability of traditional methods while combining the efficiency and precision of modern technologies. This is of great significance for shortening the breeding cycle, improving breeding efficiency, and promoting the development of the enoki mushroom industry. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a method for breeding enoki mushrooms, which solves the problems of deficiencies in existing enoki mushroom breeding methods.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a method for breeding enoki mushrooms, comprising the following steps:

[0007] S1. Collect a wide range of enoki mushroom strains from different regions, with no fewer than 50 strains, covering both wild and cultivated types;

[0008] S2. In an environment with a temperature of 20-25℃ and a relative humidity of 70%-80%, the collected strains were observed morphologically for 10 days, and strains with a cap diameter of 2-5 cm, a stipe length of 10-15 cm, and a stipe thickness of 0.2-0.5 cm were recorded.

[0009] S3. On standard culture medium, the strains were cultured at a temperature of 23±1℃, a light intensity of 500-800 lux, and a light exposure of 10-12 hours per day. Their growth rate was measured, and strains with a growth rate of 0.5-0.8 cm per day were screened out.

[0010] S4. Using artificial inoculation with pests and diseases, the strains were tested for their resistance to pests and diseases for 15 days in an environment with a temperature of 25±2℃ and a relative humidity of 85%, and strains with a pest and disease infection rate of less than 10% were selected.

[0011] S5. Select strains with a growth rate greater than 0.6 cm / day, a biological conversion rate higher than 80%, and strong resistance to diseases and pests, and hybridize them at a ratio of 1:1. The hybridization temperature is controlled at 22±1℃, and the relative humidity is maintained at 75%.

[0012] S6. The hybrid offspring were screened in an environment with a temperature of 20-23℃ and a carbon dioxide concentration of 0.1%-0.2%, and individuals with round caps, straight stems and white color were selected.

[0013] S7. The selected individuals were subjected to five consecutive generations of self-pollination purification under conditions of 18-20℃ and 60%-70% relative humidity, with each generation of self-pollination lasting 30 days.

[0014] S8. The selected superior strains are then cultured in an environment with a temperature of 15-18℃ and a relative humidity of 80%-90% for further expansion and then promoted for planting.

[0015] Preferably, in collecting samples of enoki mushroom strains from different regions, S1 ensures that the collection scope covers at least 5 different geographical areas.

[0016] Preferably, in the S2 step of morphological observation of the collected strains, strains with cap thickness in the range of 0.1-0.2 cm are recorded.

[0017] Preferably, the standard culture medium in S3 comprises: 20 g glucose, 5 g peptone, 1 g potassium dihydrogen phosphate, 0.5 g magnesium sulfate, and 15 g agar, diluted to 1 liter.

[0018] Preferably, in step S4, the inoculated pests and diseases include common brown spot pathogens and mites found in enoki mushrooms, with a pathogen concentration of 1×10⁻⁶. 6 The mite density is 5 individuals per milliliter and 5 individuals per square centimeter.

[0019] Preferably, in step S5, sterile tools are used during the hybridization operation, and the operation is performed in a sterile environment.

[0020] Preferably, in S7, the genetic stability of the strain is periodically tested during self-pollination.

[0021] This invention provides a method for breeding enoki mushrooms. It has the following beneficial effects:

[0022] 1. This invention, by extensively collecting samples of enoki mushroom strains from different regions and conducting morphological observations and growth rate measurements, can screen out strains with excellent traits. Furthermore, by artificially inoculating the strains with pests and diseases, the disease and pest resistance of the strains is tested, further ensuring that the selected enoki mushroom strains have strong disease and pest resistance, which helps to improve the yield and quality of enoki mushrooms.

[0023] 2. Through hybridization and self-pollination purification, this invention can further improve the growth rate, bioconversion rate and disease and pest resistance of enoki mushroom strains. At the same time, in the stage of large-scale cultivation and promotion, placing the finally selected superior strains in a suitable temperature and humidity environment is conducive to maintaining their excellent traits and realizing large-scale production. Detailed Implementation

[0024] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0025] Example 1:

[0026] This invention provides a method for breeding enoki mushrooms, comprising the following steps:

[0027] Sixty *Flammulina velutipes* strains were collected from five different geographical regions, including 30 wild-type and 30 cultivated-type strains. Morphological observation was conducted for 10 days at 22°C and 75% relative humidity. Strains with cap diameters of 3-4 cm, stipe lengths of 12-14 cm, stipe thicknesses of 0.3-0.4 cm, and cap thicknesses of 0.15 cm were recorded.

[0028] Then, these strains were placed on a standard culture medium prepared by adjusting the volume to 1 liter with 20 g glucose, 5 g peptone, 1 g potassium dihydrogen phosphate, 0.5 g magnesium sulfate, and 15 g agar. They were cultured at 23°C, 600 lux light intensity, and 11 hours of light per day to screen out strains with a growth rate of 0.6-0.7 cm per day.

[0029] Next, using artificial inoculation of common brown spot pathogens (concentration of 1×10⁶ CFU / mL) and mites (density of 5 mites / cm²), the strains were tested for disease and pest resistance for 15 days in an environment of 25℃ and 85% relative humidity. Strains with a disease and pest infection rate of 8% were selected. Strains with a growth rate greater than 0.65 cm / day, a biotransformation rate higher than 85%, and strong disease and pest resistance were selected and hybridized at a 1:1 ratio in a sterile environment using sterile tools. The hybridization temperature was controlled at 22℃ and the relative humidity was maintained at 75%.

[0030] Hybrid offspring were screened at 21℃ and 0.15% carbon dioxide to select individuals with rounded caps, straight stipes, and pure white color. These selected individuals underwent five consecutive generations of self-pollination purification at 19℃ and 65% relative humidity, with each generation self-pollinating for 30 days. Genetic stability of the strains was periodically monitored during self-pollination. Finally, the superior strains selected were cultured at 16℃ and 85% relative humidity for large-scale cultivation and promoted for wider distribution.

[0031] Example 2:

[0032] This invention provides a method for breeding enoki mushrooms, comprising the following steps:

[0033] Seventy *Flammulina velutipes* strains were collected from six different regions, including 35 wild-type and 35 cultivated-type strains. Morphological observation was conducted for 10 days at 20°C and 70% relative humidity, recording strains with cap diameters of 2-3 cm, stipe lengths of 10-12 cm, stipe thicknesses of 0.2-0.3 cm, and cap thicknesses of 0.1 cm.

[0034] On standard culture medium, at a temperature of 22℃, a light intensity of 500 lux, and 10 hours of light per day, strains with a growth rate of 0.5-0.6 cm per day were screened. These strains were then artificially inoculated with pests and diseases and tested for 15 days at 23℃ and 85% relative humidity, selecting strains with a pest and disease infection rate of 5%. Strains with a growth rate of 0.62 cm / day, a biotransformation rate of 82%, and strong resistance to pests and diseases were selected and hybridized at a 1:1 ratio in a sterile environment at 21℃ and 75% relative humidity.

[0035] Hybrid offspring were screened in an environment of 20℃ and 0.1% carbon dioxide concentration. After selecting superior individuals, they were purified by self-pollination for five consecutive generations at 18℃ and 60% relative humidity, with each generation lasting 30 days. Genetic stability was tested regularly. Finally, the superior strain was expanded and promoted for planting in an environment of 15℃ and 80% relative humidity.

[0036] Example 3:

[0037] This invention provides a method for breeding enoki mushrooms, comprising the following steps:

[0038] Eighty *Flammulina velutipes* strains from seven different geographical regions were collected, including 40 wild-type and 40 cultivated-type strains. Morphological observation was conducted for 10 days at 25℃ and 80% relative humidity, recording strains with cap diameters of 4-5 cm, stipe lengths of 13-15 cm, stipe thicknesses of 0.4-0.5 cm, and cap thicknesses of 0.2 cm.

[0039] Strains with a growth rate of 0.7-0.8 cm / day were selected by culturing on standard culture medium at 24℃, light intensity of 800 lux, and 12 hours of light per day. After artificial inoculation with pests and diseases and testing for 15 days at 27℃ and 85% relative humidity, strains with a pest and disease infection rate of 3% were selected. Strains with a growth rate of 0.75 cm / day, a biotransformation rate of 88%, and strong resistance to pests and diseases were selected and hybridized at a 1:1 ratio in a sterile environment at 23℃ and 75% relative humidity.

[0040] The hybrid offspring were screened in an environment of 23℃ and 0.2% carbon dioxide concentration. After selecting ideal individuals, they were purified by self-pollination for five consecutive generations at 20℃ and 70% relative humidity, with each generation lasting 30 days. Genetic stability was tested regularly during this period. Finally, the superior strains were cultured and promoted for widespread cultivation in an environment of 18℃ and 90% relative humidity.

[0041] The above embodiments enable the collection of diverse enoki mushroom strains from different regions. Through multiple rounds of meticulous screening and cultivation, hybridization, and purification under strict conditions, high-quality strains are selected. These strains exhibit rapid growth, strong resistance to diseases and pests, excellent morphology, and genetic stability, laying the foundation for the efficient and high-quality production of enoki mushrooms.

[0042] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for breeding Flammulina velutipes, characterized by, Comprise the following steps: S1. Collect a large number of different regions of the sample strain of Flammulina velutipes, the number is not less than 50, covering wild type and cultivated type; S2. In the temperature of 20-25℃, relative humidity of 70%-80% environment, the collected strains were observed for 10 days, recording the diameter of the cap range of 2-5 cm, the length of the stem is 10-15 cm and the stem thickness is 0.2-0.5 cm of the strain; S3. In the standard medium, the strain was cultured under the condition of temperature 23±1℃, light intensity 500-800 lux, light for 10-12 hours per day, the growth rate was measured, and the strain with growth rate of 0.5-0.8 cm per day was selected; S4. Using artificial inoculation of disease and insect pests, in the temperature of 25±2℃, relative humidity of 85% environment, the strain was detected for 15 days of disease and insect pests resistance, and the strain with infection rate less than 10% was selected; S5. Select the strain with growth rate greater than 0.6 cm / day, biological conversion rate higher than 80%, and strong disease and insect pests resistance, and cross at a ratio of 1:1, the cross temperature is controlled at 22±1℃, and the relative humidity is kept at 75%; S6. The selected individuals in the temperature of 20-23℃, carbon dioxide concentration of 0.1%-0.2% environment were screened, and the individuals with round cap, straight stem and white color were selected; S7. The selected individuals were self-purified under the condition of temperature 18-20℃, relative humidity 60%-70% for 5 generations, and the self-crossing time was 30 days per generation; S8. The final selected excellent strain was cultured in the temperature of 15-18℃, relative humidity of 80%-90% environment, and was popularized and planted.

2. The method for breeding Flammulina velutipes according to claim 1, characterized in that, The S1 in collecting different regions of the sample strain of Flammulina velutipes, ensures that the collection range covers at least 5 different geographical regions.

3. The method for breeding Flammulina velutipes according to claim 1, characterized in that, The S2 in the morphological observation of the collected strains, records the strain with cap thickness in the range of 0.1-0.2 cm.

4. The method for breeding Flammulina velutipes according to claim 1, characterized in that, The S3 in the standard medium includes: glucose 20 g, peptone 5 g, potassium dihydrogen phosphate 1 g, magnesium sulfate 0.5 g, agar 15 g, constant volume to 1 liter.

5. The method for breeding Flammulina velutipes according to claim 1, characterized in that, In the S4, the inoculated pests include the common brown spot pathogen and mites of the golden needle mushroom, the concentration of the pathogen is 1×10 6 per milliliter, and the density of the mites is 5 per square centimeter.

6. The method for breeding Flammulina velutipes according to claim 1, characterized in that, In the S5, sterile tools are used in the hybridization operation, and the operation is carried out in a sterile environment.

7. The method for breeding Flammulina velutipes according to claim 1, characterized in that, In the S7, the genetic stability of the strain is detected regularly during the self-crossing period.