A cultivation method of lyophyllum connatum

Abstract

The application provides a cultivation method of Lyophyllum decastes, and belongs to the technical field of edible fungi. The cultivation method of Lyophyllum decastes provided by the application comprises the following steps: inoculating the Lyophyllum decastes spores into culture material to carry out mycelium cultivation, primordium cultivation and fruiting body cultivation; the environmental conditions of the mycelium cultivation are as follows: the temperature is 19-22 DEG C, the air relative humidity is 70-75%, the carbon dioxide concentration is 800-1000 ppm, and there is no light. The application adjusts the temperature, light, humidity, carbon dioxide concentration and other factors in the stages of Lyophyllum decastes cultivation, primordium differentiation and fruiting body growth, obtains the best combination of comprehensive and coordinated control of the four factors of temperature, humidity, light and air, and provides technical support for the factory production of Lyophyllum decastes.

Description

Technical Field

[0001] This invention relates to the field of edible fungi technology, and in particular to a method for cultivating lotus leaf agaricus. Background Technology

[0002] Lyophyllum decastes (Fr.:Fr.)sing. is a very precious edible fungus with high nutritional value.

[0003] In 2005, the domestication and cultivation of *Anacarya palustris* (a type of lotus leaf) was successfully achieved in the laboratory. Fruiting bodies were obtained using both bottle and bag cultivation methods, with a biological conversion rate of 66% and a growth period of 120 days. This marked the first successful artificial cultivation of wild *Anacarya palustris* in China. Since then, after years of improvement, the artificial cultivation technology of *Anacarya palustris* has made certain progress and has been widely applied in production. In recent years, industrialized cultivation technology has rapidly emerged, while artificial cultivation technology has gradually declined. Besides the reasons related to the culture medium formulation, the control of environmental conditions is particularly important. In the artificial cultivation of *Amanita muscaria*, the control of environmental factors such as temperature, relative humidity, light intensity, and carbon dioxide concentration during mycelial growth (culture), primordia differentiation, and fruiting body growth is mainly based on single-factor control. While ventilation reduces carbon dioxide concentration, temperature can also decrease or increase due to the entry of cold or hot air, and water vapor can escape with the exhaust, thus reducing humidity. The lack of comprehensive, coordinated, and precise control of the four factors of temperature, humidity, light, and air leads to low biological conversion rate, low and unstable yield of *Amanita muscaria*, and an inability to meet the demands of industrialized production. Therefore, addressing the technical deficiencies of artificial cultivation techniques and based on the actual needs of industrialized production, a suitable environmental control condition for the industrialized production of *Amanita muscaria* is needed to provide technical support for its industrialized production. Summary of the Invention

[0004] The purpose of this invention is to provide a cultivation method for *Amanita muscaria*, which comprehensively coordinates the light intensity, air temperature, relative humidity, and carbon dioxide concentration during the mycelial growth, primordium formation, and fruiting body growth stages of *Amanita muscaria*, and precisely controls the cultivation conditions to provide technical support for the industrialized production of *Amanita muscaria*.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides a method for cultivating lotus leaf pleated cap, comprising the following steps:

[0007] The aforementioned *Agaricus rotundus* strain was inoculated into the culture medium for mycelial culture, fruiting body primordia culture, and fruiting body culture.

[0008] The environmental conditions for mycelial culture were: temperature 19–22℃, relative humidity 70–75%, carbon dioxide concentration 800–1000 ppm, and no light.

[0009] Preferably, the environmental conditions for the primordia culture of the fruiting bodies are: temperature of 13-18℃, relative humidity of air of 85-90%, carbon dioxide concentration of 550-850ppm, light intensity of 80-100lx, and light duration of 7.5-8.5h.

[0010] Preferably, the environmental conditions for fruiting body cultivation are: temperature of 16-19℃, relative humidity of 85-89%, carbon dioxide concentration of 850-1100ppm, light intensity of 80-100lx, and 12-hour light-dark alternation.

[0011] Preferably, the culture medium is composed of the following raw materials in the indicated mass fractions: 35-45% corn cobs, 18-22% cottonseed hulls, 3-7% cornmeal, 10-30% king oyster mushroom spawn, 10-20% wheat bran, 1-5% soybean meal, and 1-3% gypsum.

[0012] This invention achieves the optimal combination of temperature, humidity, light, and air quality through adjusting factors such as temperature, light, humidity, and carbon dioxide concentration during the stages of mycorrhizal growth, primordia differentiation, and fruiting body development of lotus leaf agaric. This provides technical support for the industrialized production of lotus leaf agaric. Detailed Implementation

[0013] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0014] Example 1

[0015] A method for cultivating lotus leaf umbelliferous foliage includes the following steps:

[0016] The liquid spawn of *Pleurotus ostreatus*, which had been incubated for 8 days, was inoculated into the culture medium. The culture medium consisted of the following ingredients in the indicated mass fractions: 35% corn cob, 20% cottonseed hulls, 5% cornmeal, 20% *Pleurotus ostreatus* spawn, 15% wheat bran, 3% soybean meal, and 2% gypsum. Water was added to bring the moisture content to 67%. Mycelial culture was carried out on this culture medium under the following environmental conditions: temperature range of 19–22°C, relative humidity of 73%, carbon dioxide concentration of 900 ppm, and no light.

[0017] After the mycelium has fully colonized and completed physiological maturation (i.e., the mycelium has completely filled the culture bottle), continue cultivation for 8 days, then proceed to the primordium cultivation and management stage. Adjust the environmental conditions as follows: maintain 18℃ from 6:00 AM to 6:00 PM, and maintain 13℃ from 6:00 PM to 6:00 AM the following day, with a diurnal temperature difference of 5℃. Temperature fluctuations should be gentle, with a 1℃ increase or decrease every hour, reaching the target within 5 hours. Maintain a relative humidity of 87% (constant humidity for 24 hours), alternating between 12 hours of light (light intensity 90 lx) and 12 hours of darkness, and a CO2 concentration of 700 ppm for 10 days. When primordium differentiation reaches 95% or more in 95% of the culture bottles, proceed to the fruiting body growth stage.

[0018] The environmental conditions for fruiting body growth period cultivation are: temperature range of 16-19℃, relative humidity of 87%, carbon dioxide concentration of 1000ppm, light intensity of 90lx, and 12-hour light-dark alternation.

[0019] Comparative Example 1

[0020] To investigate the environmental conditions for the growth of *Amanita muscaria* mycelium, inoculated bottles were placed in incubation chambers with comprehensive conditions I, II, and III as shown in Table 1. Each incubation chamber contained 1000 bottles, for a total of 3000 bottles. The incubation process followed the method described in Example 1. Thirty bottles were randomly selected from each comprehensive condition. The number of bottles contaminated, mycelial growth, mycelial vigor, and time to full colonization were periodically observed and recorded. The contamination rate, mycelial growth rate, and average time to full colonization were statistically analyzed. Through significance testing, the suitable comprehensive conditions for mycelial growth were determined. Comprehensive condition II corresponds to the incubation conditions in Example 1.

[0021] Table 1. Environmental conditions for the growth of *Agropyron cristatum* mycelium on lotus leaves.

[0022]

[0023] The results are shown in Table 2. The growth rate of *Amanita muscaria* mycelium under comprehensive conditions II (temperature 19–22℃, relative humidity 73%, carbon dioxide concentration 900 ppm, light intensity 0 Lx) (3.93 mm / d) was significantly higher than that under comprehensive conditions I (2.85 mm / d) and comprehensive conditions III (3.2 mm / d); the contamination rate (5.6%) was significantly lower than that under comprehensive conditions I (28.5%) and comprehensive conditions III (10.3%); and the number of days for mycelium to fully colonize the culture bottle (50.90 days) was significantly shorter than that under the other two comprehensive conditions.

[0024] Table 2. Results of the study on suitable growth conditions for *Agropyron cristatum* mycelium on lotus leaves.

[0025]

[0026] Comparative Example 2

[0027] To determine the optimal comprehensive conditions (temperature, temperature difference, relative humidity, light, and carbon dioxide concentration) for primordia differentiation of *Amanita muscaria* fruiting bodies, 3000 bottles with fully colonized mycelia and completed physiological after-ripening (8 days of continued cultivation after mycelial colonization) under condition 2 of Comparative Example 1 were randomly divided into three groups of 1000 bottles each, placed in environments with comprehensive conditions 1, 2, and 3 (as shown in Table 3), respectively. The cultivation procedures were performed according to Example 1. Thirty bottles were randomly selected from each comprehensive condition, and the number of primordia formed and the number of mature mushrooms were observed and recorded. Significance tests were performed on the data to determine the optimal comprehensive conditions for primordia formation of *Amanita muscaria* fruiting bodies. Comprehensive condition III refers to the cultivation conditions of Example 1.

[0028] Table 3. Comprehensive conditions for the differentiation of primordia of lotus leaf-folded umbelliferous fruiting bodies

[0029]

[0030] The experimental results are shown in Table 4. Under environmental conditions I (temperature 21–25℃, humidity 87%, carbon dioxide concentration 500ppm, light intensity 70Lx, temperature difference 3–5℃), the mycelium of *Amanita muscaria* could not differentiate, and the number of primordia formed was zero. Under environmental conditions III (temperature 13–18℃, humidity 87%, carbon dioxide concentration 600ppm, light intensity 90Lx, temperature difference 3–5℃), the number of mushroom primordia formed by *Amanita muscaria* was significantly higher than that under environmental conditions II (temperature 15–20℃, humidity 93%, carbon dioxide concentration 600ppm, light intensity 50Lx, temperature difference 3–5℃).

[0031] Table 4. Results of research on suitable environmental conditions for the differentiation of *Apis lutea* primordia.

[0032] Comprehensive conditions Primordial differentiation number Primary basal mushroom growth rate Ⅰ 0 0 Ⅱ 22.75 13.5 Ⅲ 27.45 18.5**

[0033] Comparative Example 3

[0034] To determine the optimal comprehensive conditions (temperature, relative humidity, light intensity, light duration, and carbon dioxide concentration) for the growth of *Umbelliferae rubra* fruiting bodies, 3000 vials that had completed primordia differentiation (primordia the size of rice grains) under condition 3 in Comparative Example 2 were randomly divided into three groups of 1000 vials each. These were placed in environments with comprehensive conditions 1, 2, and 3 (as shown in Table 5), respectively, and the cultivation procedures were performed according to Example 1. Twenty vials were randomly selected, and the growth of the fruiting bodies (number of primordia forming mushrooms, stipe length, diameter, weight, and color; cap diameter, thickness, color, and weight) were observed and recorded. The significance of the data was determined to identify the optimal comprehensive conditions for the growth of *Umbelliferae rubra* fruiting bodies. Comprehensive condition III refers to the cultivation conditions of Example 1.

[0035] Table 5 Suitable environmental conditions for the growth and development of lotus leaf pleated umbellets

[0036]

[0037] The experimental results are shown in Table 6. After the formation of the primordia of *Anacarya paliurus*, the biological conversion rate of *Anacarya paliurus* under environmental condition III (temperature 16-19℃, temperature difference 2-3℃, humidity 87%, carbon dioxide concentration 1000ppm, light intensity 90Lx, 12 hours of light-dark alternation) reached 94.5%, and the fresh weight per bottle reached 378.47g. This was significantly higher than the biological conversion rate (88.2%) and fresh weight per bottle (353.20g) under environmental condition II, and extremely significantly higher than the biological conversion rate (66.4%) and fresh weight per bottle (277.64g) under environmental condition I.

[0038] Table 6. Results of the study on suitable environmental conditions for the growth of lotus leaf-puffed umbelliferous fruiting bodies.

[0039]

[0040] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for cultivating *Aureobasidium alatum*, characterized in that, Includes the following steps: The aforementioned *Agaricus rotundus* strain was inoculated into the culture medium for mycelial culture, fruiting body primordia culture, and fruiting body culture. The environmental conditions for mycelial culture were: temperature 19-22℃, relative humidity 73%, carbon dioxide concentration 900ppm, and no light. The environmental conditions for the primordia culture of the fruiting bodies are as follows: the temperature is 13~18℃, maintained at 18℃ from 6:00 am to 6:00 pm, and maintained at 13℃ from 6:00 pm to 6:00 am the next day, with a temperature fluctuation of 1℃ per hour, and the standard is reached in 5 hours. The relative humidity of the air is 87%, the carbon dioxide concentration is 600 ppm, the light intensity is 90 lx, and the light exposure time is 8 hours. The environmental conditions for fruiting body cultivation are: temperature 16~19℃, relative humidity 87%, carbon dioxide concentration 1000ppm, light intensity 90lx, and 12-hour light-dark alternation.

2. The cultivation method according to claim 1, characterized in that, The culture medium is composed of the following raw materials in the indicated mass fractions: corn cob 35-45%, cottonseed hull 18-22%, cornmeal 3-7%, king oyster mushroom spawn 10-30%, wheat bran 10-20%, soybean meal 1-5%, and gypsum 1-3%.

Images