Dictyophora rubrovolvata bud in-vitro differentiation fruiting method

Abstract

The invention discloses a dictyophora rubrovolvata bud in-vitro differentiation fruiting method. The method comprises the following steps: harvesting 7-9% of mature dictyophora rubrovolvata buds; the method specifically comprises the following steps: arranging a microporous ceramic plate on a bottom plate of an incubator, forming a culture layer and a liquid storage layer in the incubator, laying a layer of pine needles on the microporous ceramic plate, placing collected mature dictyophora rubrovolvata buds of 7-9% on the pine needles, and keeping the temperature of a culture environment at 21-27 DEG C; the liquid storage layer is regularly filled with a culture nutrient solution, so that the roots of dictyophora rubrovolvata buds and the laid pine needles discontinuously make contact with the culture nutrient solution; a multispectral imager is arranged in the center of the top of the incubator, and whether buds of dictyophora rubrovolvata break shells, stipes of dictyophora rubrovolvata extend and whether dictyophora rubrovolvata can be picked or not are obtained in real time according to monitored data. According to the method, nutrient and water supply required by growth of dictyophora rubrovolvata can be controlled, and growth in different stages can be monitored in real time, so that whole-process accurate management of growth of dictyophora rubrovolvata is realized, the fruiting rate is increased, and large-scale production is facilitated.

Description

Technical Field

[0001] The present invention relates to the technical field of edible mushroom production. More specifically, the present invention relates to a method for in vitro differentiation and fruiting of Dictyophora rubrovolvata mushroom buds. Background Art

[0002] Dictyophora rubrovolvata is a unique edible mushroom variety in China, with unique biological characteristics. The development of the fruiting body of this variety shows significant time-effect characteristics. It only takes 4 - 6 hours from the cracking of the mushroom egg to full maturity, and this process mostly occurs in the early morning. This rapid development characteristic poses strict requirements for the harvesting work: if not harvested in time, the spores of the mature fruiting body will autolyze within a short time, and the mushroom body will shrink due to water loss, color deterioration, and texture change due to aging, resulting in a significant decline in edible quality and commercial value. The fresh product shows significant changes in nutritional components within 2 - 3 hours under normal temperature conditions.

[0003] Due to its small size, regular shape, and strong compressive resistance, the mushroom buds of Dictyophora rubrovolvata have significant advantages in storage and transportation. If the process of developing from mushroom buds to mature fruiting bodies can be achieved at the consumer end, it will effectively solve the following pain points faced by the current Dictyophora rubrovolvata industry: high logistics costs and unstable product quality caused by special cultivation environments, scattered production areas, immediate harvesting requirements, and strict fresh product preservation conditions. However, the formation of the mushroom cap and the elongation and unfolding of the stalk skirt are complex physiological processes regulated by multiple factors. If the conditions are not properly controlled, it may lead to premature or abnormal development of the mushroom buds, or even termination of development, seriously affecting the commerciality. Therefore, developing a reliable in vitro differentiation and fruiting technology for Dictyophora rubrovolvata is of great significance for the healthy and sustainable development of the industry. Summary of the Invention

[0004] The present invention provides a method for in vitro differentiation and fruiting of Dictyophora rubrovolvata mushroom buds, which can control the supply of nutrients and water required for the growth of Dictyophora rubrovolvata, and monitor the growth at different stages in real time, so as to achieve precise management of the whole growth process of Dictyophora rubrovolvata, promote its healthy growth and development, improve the fruiting rate and success rate, and facilitate large-scale production.

[0005] To achieve these and other advantages of the present invention, a method for in vitro differentiation and fruiting of Dictyophora rubrovolvata mushroom buds is provided, including: Harvesting 7 - 9 mature Dictyophora rubrovolvata mushroom buds at the Dictyophora rubrovolvata planting site; Culturing the 7 - 9 mature Dictyophora rubrovolvata mushroom buds, specifically: setting a microporous ceramic plate on the bottom plate of the incubator, so that the incubator forms a culture layer on the upper part and a liquid storage layer on the lower part, laying a layer of pine needles on the microporous ceramic plate, placing the collected 7 - 9 mature Dictyophora rubrovolvata mushroom buds on the pine needles, and maintaining the temperature of the culture environment at 21 - 27°C; Regularly fill the liquid storage layer with a culture nutrient solution, so that the roots of the Dictyophora rubrovolvata mushroom buds and the laid pine needles intermittently contact the culture nutrient solution, supplementing nutrients and moisture to the Dictyophora rubrovolvata mushroom buds; A multispectral imager is arranged at the central position on the top of the incubator to monitor the growth of Dictyophora rubrovolvata at different stages through the multispectral imager, and to obtain in real time whether the mushroom buds of Dictyophora rubrovolvata have broken their shells, whether the stipes have elongated, and whether they can be picked through the monitored data.

[0006] Preferably, the components of the culture nutrient solution at different growth stages of Dictyophora rubrovolvata are as follows: Before the shell-breaking stage, yeast extract 0.08%-0.12%, glucose 0.15%-0.25%, magnesium sulfate 0.02%-0.04%, potassium dihydrogen phosphate 0.03%-0.05%, vitamin B1 0.0008%-0.0012%, pH 6.0-6.5, and the balance is water; During the shell-breaking stage, an additional 0.04%-0.06% of potassium dihydrogen phosphate and 0.08%-0.12% of L-arginine are added; During the stipe elongation stage, glucose is increased to 0.25%-0.35%, and zinc sulfate is 0.004%-0.006%; Before the harvesting stage, L-glutamine is 0.1%-0.2%, and glucose is decreased to 0.08%-0.12%.

[0007] Preferably, the pine needles laid on the microporous ceramic plate need to be pretreated, and the pretreatment method is as follows: A. Sterilize the pine needles in steam at 80-100°C for 30-60 minutes; B. Immerse the sterilized pine needles in the pine needle nutrient solution for 6-12 hours. The components of the pine needle nutrient solution are 0.05%-0.1% yeast extract, 0.1%-0.3% glucose, 0.01%-0.05% magnesium sulfate, and 0.02%-0.06% potassium dihydrogen phosphate, and the balance is water.

[0008] Preferably, the specific implementation method of regularly filling the liquid storage layer with the culture nutrient solution is as follows: The vertical distance between the microporous ceramic plate and the bottom plate of the incubator is 3-5 cm, and a liquid level sensor is arranged on the side wall of the liquid storage layer; A liquid inlet and a liquid outlet are arranged on the side wall of the liquid storage layer. The liquid inlet is connected to an external container through a first pipeline, and the liquid outlet is connected to the external container through a second pipeline. A micro pump and a first solenoid valve are arranged in the first pipeline, a second solenoid valve is arranged in the second pipeline, and the height of the liquid storage layer is higher than the height of the external container; Cultivation nutrient solution rising stage: Open the first solenoid valve and the micro pump, close the second solenoid valve, and inject the cultivation nutrient solution into the liquid storage layer. When the liquid level sensor detects that the liquid level has risen to contact the lower surface of the microporous ceramic plate, close the first solenoid valve and the micro pump, stop the liquid injection, and then slowly moisten the pine needles through the capillary action of the microporous ceramic plate without submerging the mushroom buds. Cultivation nutrient solution falling stage: Open the second solenoid valve and drain the cultivation nutrient solution in the liquid storage layer into the external container. Among them, it circulates 1 - 3 times a day, each time the liquid level is maintained for 5 - 20 minutes. When soaking, the height of the nutrient solution liquid level is 1 / 3 - 1 / 2 of the thickness of the pine needles, avoiding completely submerging the mushroom buds. Among them, the incubator is an incubator with an open top.

[0009] Preferably, a 100 - mesh filter screen is provided at the inlet end of the first pipeline. Sterile air is continuously introduced into the cultivation nutrient solution during the infiltration stage, and the ventilation holes are located 1 cm from the bottom plate on the side wall of the liquid storage layer.

[0010] Preferably, the multispectral imager is used to monitor the growth of Dictyophora rubrovolvata at different stages. The specific process is as follows: The multispectral imager is 30 - 50 cm away from the mushroom bud, and the field of view angle covers the entire incubator. Multispectral images during the growth process of the mushroom bud are obtained through the multispectral imager and analyzed by an analysis module, including: Obtain visible light band images to analyze the color change of the outer skin of the mushroom bud to determine whether it has broken its shell. Obtain near - infrared band images to monitor the elongation process of the mushroom stalk to determine whether the mushroom stalk has elongated. Obtain short - wave infrared band images to detect the mushroom skirt expansion index to determine whether it can be harvested.

[0011] Preferably, the multispectral images during the growth process of the mushroom bud are obtained through the multispectral imager and analyzed by an analysis module. The specific analysis process is as follows: Extract the reflectance data of the volva area from the visible light band images, calculate the red edge index REI. When the red light band is 600 - 700 nm and the REI decline rate reaches 8% - 15%, it is determined that the outer skin of the mushroom bud begins to crack, that is, it begins to break its shell. According to the near - infrared band images, calculate the normalized difference water index NDWI. When the NDWI value continuously rises, it indicates that the mushroom stalk enters the rapid growth period. Calculate the mushroom skirt expansion index SVI from the short - wave infrared band images. When SVI > 0.3 and the drooping angle of the mushroom skirt > 120°, it indicates that it can be harvested.

[0012] Preferably, it also includes adjusting the environmental parameters, specifically: When the decline rate of the red-edge index REI reaches 8%-15% and it is determined that the outer skin of the mushroom bud begins to crack, lower the temperature in the incubator by 2 - 3°C, and at the same time adjust the light intensity to 200 - 300 lux; Increase the air circulation rate in the incubator to 0.5 - 0.8 m / s to ensure sufficient oxygen supply and promote the growth of the mushroom stalk.

[0013] Preferably, if the shell-breaking is delayed and the REI decline rate <0.5% / h lasts for 24 hours and the NDWI does not increase, then: Gently scratch the top of the mushroom bud with a sterile needle tip, or slightly vibrate the surface of the mushroom bud through ultrasonic waves; Synchronously increase the humidity of the culture layer to 90%.

[0014] Preferably, the pore diameter of the microporous ceramic plate is 20-50 μm, the porosity is 45-55%, the laying density of pine needles is 0.15-0.25 g / cm³, and the thickness of the pine needle layer is 3-5 cm.

[0015] The present invention has at least the following beneficial effects: First, by setting a microporous ceramic plate in the incubator to form a culture layer and a liquid storage layer, the supply of nutrients and water required for the growth of Dictyophora rubrovolvata can be accurately controlled. Different components of culture nutrient solutions are used in different growth stages to meet the specific nutritional requirements of Dictyophora rubrovolvata in various stages such as the pre-shell-breaking stage, shell-breaking stage, mushroom stalk elongation stage, and pre-harvest stage, which helps its healthy growth and development.

[0016] Second, by using a multi-spectral imager set at the center position of the top of the incubator, the growth conditions of Dictyophora rubrovolvata at different stages can be monitored in real time. By analyzing images in different bands, such as visible light band images for analyzing the color change of the outer skin of the mushroom bud, near-infrared band images for monitoring the elongation process of the mushroom stalk, and short-wave infrared band images for detecting the expansion index of the mushroom skirt, it is possible to accurately judge whether the mushroom bud has broken its shell, whether the mushroom stalk has elongated, and whether it can be harvested, providing a scientific basis for precise management and harvesting.

[0017] Third, the pre-treatment of the laid pine needles, including steam sterilization and soaking in pine needle nutrient solution, provides a more suitable growth substrate for the mushroom buds of Dictyophora rubrovolvata. In addition, during the cultivation process, the environmental parameters are adjusted according to the growth state of Dictyophora rubrovolvata, such as lowering the temperature, adjusting the light intensity when the outer skin of the mushroom bud begins to crack, and increasing the air circulation rate to ensure oxygen supply, which helps to promote the growth and development of Dictyophora rubrovolvata. For the case of delayed shell-breaking, measures such as gently scratching the top of the mushroom bud with a sterile needle tip or slightly vibrating the surface of the mushroom bud through ultrasonic waves and increasing the humidity of the culture layer can effectively solve the shell-breaking problem and improve the mushroom yield.

[0018] Fourth, the circulating supply mode of the culture nutrient solution, as well as the precise control and monitoring of the culture environment parameters, make the growth process of Dictyophora rubrovolvata more stable and controllable, reduce the blindness of manual intervention, improve the culture efficiency and success rate, and are conducive to the large-scale culture and production of Dictyophora rubrovolvata.

[0019] Other advantages, objectives and features of the present invention will be partially reflected by the following description, and partially will also be understood by those skilled in the art through the research and practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a schematic flow chart of the method for in vitro differentiation and fruiting of Dictyophora rubrovolvata mushroom buds of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0021] The following further describes the present invention in detail with reference to the drawings, so that those skilled in the art can implement it according to the description in the specification.

[0022] It should be understood that the terms such as "having", "comprising" and "including" used herein do not exclude the presence or addition of one or more other elements or their combinations.

[0023] It should be noted that the experimental methods described in the following embodiments are all conventional methods unless otherwise specified, and the reagents and materials can be obtained from commercial channels unless otherwise specified; in the description of the present invention, it should be noted that unless otherwise clearly defined and limited, the terms "installation", "connection" and "setting" should be understood in a broad sense. For example, it can be fixedly connected and set, or detachably connected and set, or integrally connected and set. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific situations. The orientation or positional relationship indicated by the terms "horizontal", "longitudinal", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings, and is only for the convenience of describing the present invention and simplifying the description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation to the present invention.

[0024] As Figure 1 shown, the present invention provides a method for in vitro differentiation and fruiting of Dictyophora rubrovolvata mushroom buds, including: S1. Harvest Dictyophora rubrovolvata mushroom buds at 7-9 maturity at the Dictyophora rubrovolvata planting site.

[0025] Among them, the 7-9 mature mushroom buds are harvested at the red puffer bamboo mushroom planting site. The mushroom buds are commonly known as mushroom eggs, and their characteristics are as follows: The 7-9 mature mushroom buds are oval in shape, with a raised and hard top, a water content of not less than 70%, the mushroom buds are intact, and there are no diseases. The harvesting method is as follows: Use a small knife or scissors to cut off the whole mushroom bud or pick off the whole mushroom bud by hand, and then place it in an open container with pine needles spread at the bottom, with the root facing down and the top facing up. When stacking multiple layers, pine needles are used to separate the top and the root of the mushroom bud. The container is open and not sealed. The harvested mushroom buds are placed in a transport vehicle, with the temperature controlled below 20°C and the humidity controlled below 80%. The temperature can be between 20°C and 4°C. The lower the temperature, the longer the storage time. The harvested mushroom buds are transported to the cultivation site.

[0026] S2. Cultivate the 7-9 mature red puffer bamboo mushroom buds. Specifically, a microporous ceramic plate is arranged on the bottom plate of the incubator, so that the incubator forms a cultivation layer on the upper part and a liquid storage layer on the lower part. A layer of pine needles is laid on the microporous ceramic plate, and the harvested 7-9 mature red puffer bamboo mushroom buds are placed on the pine needles, and the temperature of the cultivation environment is maintained at 21-27°C.

[0027] Among them, the top of the incubator is open. The pore size of the microporous ceramic plate is 20-50μm, the porosity is 45-55%, the laying density of the pine needles is 0.15-0.25g / cm³, and the thickness of the pine needle layer is 3-5cm. The vertical distance between the microporous ceramic plate and the bottom plate of the incubator is 3-5cm. A liquid level sensor is arranged on the side wall of the liquid storage layer to monitor the height of the cultivation nutrient solution in the liquid storage layer.

[0028] In the above steps, when culturing the 7 - 9 mature Dictyophora indusiata fruiting bodies, first select a suitable incubator with an open - top structure. The incubator body can be made of transparent plastic or glass. Place a microporous ceramic plate on the bottom plate of the incubator. Exemplarily, the pore diameters of the microporous ceramic plate can be selected from 20μm, 30μm, 40μm, 50μm, etc., and the porosity can be 45%, 50%, 55%, etc. Install it at a vertical distance of 3 - 5 cm from the bottom plate of the incubator. Preferably, it can be installed at 3 cm, dividing the incubator into an upper culture layer and a lower liquid storage layer. In the process of laying pine needles, select the needles of common local pine tree varieties and lay a layer of pine needles with a thickness of 3 - 5 cm on the microporous ceramic plate at a laying density of 0.15 - 0.25 g / cm³. Exemplarily, lay a layer of pine needles with a thickness of 4 cm at a laying density of 0.2 g / cm³. After completing the laying of pine needles, carefully place the collected 7 - 9 mature Dictyophora indusiata fruiting bodies on the pine needles. During the whole culture process, with the help of an air conditioner or a constant - temperature device, maintain the culture environment temperature between 21 - 27°C. Install a liquid - level sensor at a suitable position on the side wall of the liquid storage layer. A common contact - type liquid - level sensor can be selected to monitor the height of the culture nutrient solution in the liquid storage layer in real time. The data monitored by the liquid - level sensor is transmitted to the control center in real time.

[0029] Therefore, in the above steps, the open - top of the open - top incubator is conducive to air circulation, providing sufficient oxygen for the growth of Dictyophora indusiata. The microporous ceramic plate with specific pore diameters and porosity can not only effectively separate the culture layer and the liquid storage layer, but also enable the culture nutrient solution in the liquid storage layer to slowly penetrate into the culture layer through capillary action, providing stable and appropriate moisture and nutrients for the Dictyophora indusiata fruiting bodies. The pine needles laid at a certain density and thickness simulate the natural growth environment of Dictyophora indusiata, providing good support and a natural - like growth substrate for the fruiting bodies. The precisely controlled culture environment temperature helps the Dictyophora indusiata fruiting bodies grow healthily, improving the success rate of culture and the quality of mushroom production.

[0030] S3. Regularly fill the liquid storage layer with the culture nutrient solution, so that the roots of the Dictyophora indusiata fruiting bodies and the laid pine needles intermittently contact the culture nutrient solution, supplementing nutrients and moisture for the Dictyophora indusiata fruiting bodies. Among them, the thickness of the pine needles immersed in the culture nutrient solution is 1 / 3 - 1 / 2 of the thickness of the pine needles.

[0031] Among them, the specific implementation method of regularly filling the liquid storage layer with the culture nutrient solution is as follows: S301. There are a liquid inlet and a liquid outlet on the side wall of the liquid storage layer. The liquid inlet is connected to an external container through a first pipeline, and the liquid outlet is connected to the external container through a second pipeline. A micro - pump and a first solenoid valve are arranged in the first pipeline, and a second solenoid valve is arranged in the second pipeline. And the height of the liquid storage layer is higher than the height of the external container; S302. Cultivation nutrient solution rising stage: Open the first solenoid valve and the micro pump, close the second solenoid valve, and inject the cultivation nutrient solution into the liquid storage layer. When the liquid level sensor detects that the liquid level rises to contact the lower surface of the microporous ceramic plate, close the first solenoid valve and the micro pump, stop the liquid injection, and then slowly moisten the pine needles through the capillary action of the microporous ceramic plate without submerging the mushroom buds. S303. Cultivation nutrient solution descending stage: Open the second solenoid valve and discharge the cultivation nutrient solution in the liquid storage layer into the external container. Among them, it circulates 1 - 3 times a day, and each time the liquid level is maintained for 5 - 20 minutes. When soaking, the height of the nutrient solution liquid level is 1 / 3 - 1 / 2 of the thickness of the pine needles, avoiding completely submerging the mushroom buds.

[0032] In the above steps, the liquid inlet and liquid outlet on the side wall of the liquid storage layer can be common plastic material interfaces, and the specifications only need to be adapted to the connecting pipelines. The first pipeline and the second pipeline can be PVC plastic hoses resistant to acid and alkali corrosion, with a pipe diameter of, for example, 8 mm or 10 mm, to ensure the smooth transportation of the culture nutrient solution. The micro pump can be a small peristaltic pump on the market, as long as it can achieve the pumping function. The first solenoid valve and the second solenoid valve can be normally closed solenoid valves, which open when powered on and close when powered off, to control the on and off of the culture nutrient solution in the pipeline. The liquid level sensor can be a contact liquid level sensor with higher precision, which can accurately sense the change of the liquid level height. The first solenoid valve, the second solenoid valve and the micro pump are all electrically connected to the control center. During specific implementation, the liquid inlet and liquid outlet are installed at appropriate positions on the side wall of the liquid storage layer. One end of the first pipeline is connected to the liquid inlet, and the other end is connected to an external container filled with the culture nutrient solution. The micro pump and the first solenoid valve are successively installed near the liquid inlet of the first pipeline. One end of the second pipeline is connected to the liquid outlet, and the other end is connected back to the external container. The first solenoid valve and the micro pump are opened through the control center, and the micro pump starts to pump the culture nutrient solution in the external container into the liquid storage layer through the first pipeline. As the liquid level rises, the liquid level sensor monitors in real time. When the liquid level sensor detects that the liquid level rises to just contact the lower surface of the microporous ceramic plate, for example, if the microporous ceramic plate is 3 cm away from the bottom of the liquid storage layer, the liquid level sensor feeds back that the liquid level reaches 3 cm at this time, and the control center immediately closes the first solenoid valve and the micro pump to stop the liquid injection. At this time, due to the capillary action of the microporous ceramic plate, the nutrient solution slowly penetrates upward and gradually wets the pine needles, but does not immerse the fruiting bodies of Dictyophora rubrovolvata. Assuming that the thickness of the pine needle layer is 5 cm, the wetted thickness is controlled within 1.7 - 2.5 cm (i.e., 1 / 3 - 1 / 2 of the pine needle thickness). After a set time, for example, 15 minutes, and each time the liquid level is maintained within the range of 5 - 20 minutes, the control center opens the second solenoid valve. Since the height of the liquid storage layer is higher than that of the external container, under the action of gravity, the culture nutrient solution in the liquid storage layer is drained back to the external container through the second pipeline, completing one cycle. One to three such cycle operations can be carried out daily according to the setting. For example, the first cycle is carried out at 8:00 in the morning, the second cycle is carried out at 12:00 noon, and the third cycle is carried out at 4:00 in the afternoon.

[0033] By intermittently allowing the pine needles and the roots of the fruiting bodies to contact the nutrient solution, it avoids the possible oxygen deficiency or other growth problems that the fruiting bodies may cause due to being overly immersed in the nutrient solution. At the same time, it can accurately provide the nutrients and moisture required for the growth of the fruiting bodies, meet their growth needs, and promote healthy growth. The culture nutrient solution is recycled between the liquid storage layer and the external container, reducing the waste of the nutrient solution, improving the utilization rate of the nutrient solution, and reducing the culture cost. By using devices such as liquid level sensors, solenoid valves and micro pumps, the automatic control of the nutrient solution supply process is realized, which is convenient for the operator to flexibly adjust according to the growth situation of Dictyophora rubrovolvata, improving the controllability and management efficiency of the culture process.

[0034] S4. A multispectral imager is set at the central position on the top of the incubator to monitor the growth of *Dictyophora rubrovolvata* at different stages, and whether the primordium of *Dictyophora rubrovolvata* breaks its shell, whether the stipe elongates, and whether it can be picked are obtained in real time through the monitored data.

[0035] Among them, the process of using the multispectral imager to monitor the growth of *Dictyophora rubrovolvata* at different stages is as follows: A. The multispectral imager is 30 - 50 cm away from the primordium, and the field of view angle covers the entire incubator; B. Multispectral images during the growth of the primordium are obtained by the multispectral imager and analyzed by an analysis module, including: Obtaining visible light band images to analyze the color change of the primordium's outer skin to judge whether it breaks its shell; Obtaining near-infrared band images to monitor the elongation process of the stipe to judge whether the stipe elongates; Obtaining short-wave infrared band images to detect the stipe expansion index to judge whether it can be collected.

[0036] Specifically, the process of obtaining multispectral images during the growth of the primordium by the multispectral imager and analyzing them by the analysis module is as follows: S401. Reflectance data of the volva region are extracted from the visible light band images, and the red edge index REI is calculated. When the red light band is 600 - 700 nm and the decline rate of REI reaches 8% - 15%, it is determined that the outer skin of the primordium begins to rupture, that is, it begins to break its shell; S402. According to the near-infrared band images, the normalized difference water index NDWI is calculated. When the value of NDWI continues to rise, it indicates that the stipe enters the rapid growth period; S403. From the short-wave infrared band images, the stipe expansion index SVI is calculated. When SVI > 0.3 and the stipe drooping angle > 120°, it indicates that it can be collected.

[0037] In the above steps, for a multi-spectral scanner, the commonly available Specim IQ multi-spectral imager on the market can be used. It has high resolution and a relatively wide spectral range, and can meet the acquisition requirements of images of different bands of Dictyophora rubrovolvata. The multi-spectral imager acquires images in the visible light (400 - 700 nm), near-infrared (700 - 1300 nm), and short-wave infrared (1300 - 2500 nm) bands. The multi-spectral imager is installed at the center position on the top of the incubator and fixed by an adjustable bracket. The adjustable bracket adopts existing technology, as long as it can achieve this function, to ensure that the imager lens is vertically downward and aligned with the mushroom bud. Adjust the distance between the multi-spectral imager and the mushroom bud to ensure that the field of view angle can completely cover the entire incubator. Use the data analysis software supporting the multi-spectral imager. This software has functions of image analysis and related index calculation. Specifically, during the cultivation process of Dictyophora rubrovolvata, start the multi-spectral imager and set the acquisition frequency. For example, acquire multi-spectral images once every 1 - 2 hours. The multi-spectral imager sequentially obtains images in the visible light band, near-infrared band, and short-wave infrared band according to the set parameters. The multi-spectral imager is installed at the center position on the top, 40 cm away from the mushroom bud, and the field of view angle can clearly cover the growth area of Dictyophora rubrovolvata in the entire incubator. The size of the incubator is not specifically limited in this embodiment and can be specifically set according to the actual situation and cultivation scale. For the visible light band image, extract the reflectance data of the volva area and calculate the red edge index REI. The calculation method is: , where R 600 is the reflectance in the 600 nm red light band, and R 700 is the reflectance in the 700 nm red light band. When the decline rate of REI reaches 8% - 15%, it indicates that the outer skin of the mushroom bud begins to rupture (hatch). For the near-infrared band image, calculate the normalized difference water index NDWI to reflect the change in the water content of the stipe: , where R 860 is the reflectance in the 860 nm near-infrared band, and R 1240 is the reflectance in the 1240 nm short-wave infrared band. When calculating NDWI, 1240 nm and 860 nm are the corresponding and appropriate bands in the short-wave infrared band and the near-infrared band. When NDWI continuously rises, it indicates that the stipe enters the rapid growth period. For the short-wave infrared band image, calculate the stipe skirt unfolding index SVI to evaluate the degree of stipe skirt unfolding: First, perform preprocessing such as image enhancement and threshold segmentation on the short-wave infrared image to segment the stipe skirt area, and then calculate the area A and perimeter P of the stipe skirt area, , when SVI > 0.3 and the stipe skirt drooping angle > 120°, it indicates that it can be collected. For the stipe skirt drooping angle, the calculation process is: Identify the top center point (A) and the outermost drooping point (B) of the stipe skirt, calculate the stipe skirt support point (C) (the connection point between the stipe and the volva), and the vector method can be used to calculate the included angle between vector CA (from the support point to the top) and vector CB (from the support point to the drooping point) : If > 120°, it indicates that the mushroom skirt is fully unfolded and meets the picking standard. By analyzing images of different bands and calculating relevant indices, it is possible to accurately judge whether the Dictyophora rubrovolvata bud has broken its shell, whether the stipe has elongated, and whether it meets the picking standard. This precise judgment provides a scientific basis for management decisions during the cultivation process. For example, during the rapid growth period of the stipe, the nutrient supply and environmental parameters can be adjusted in a timely manner to improve the quality and yield of Dictyophora rubrovolvata. The multispectral imager monitors from the top of the incubator without contacting Dictyophora rubrovolvata, avoiding interference with its growth process, ensuring that Dictyophora rubrovolvata grows in a natural state, and improving the accuracy and reliability of the monitoring results.

[0038] In one specific embodiment, the components of the culture nutrient solution at different growth stages of Dictyophora rubrovolvata are as follows: Before shell breaking, yeast extract 0.08% - 0.12%, glucose 0.15% - 0.25%, magnesium sulfate 0.02% - 0.04%, potassium dihydrogen phosphate 0.03% - 0.05%, vitamin B1 0.0008% - 0.0012%, pH 6.0 - 6.5, with the balance being water; During shell breaking, an additional 0.04% - 0.06% of potassium dihydrogen phosphate and 0.08% - 0.12% of L - arginine are added; During the stipe elongation period, the glucose is increased to 0.25% - 0.35%, and zinc sulfate is 0.004% - 0.006%; In the pre - harvest period (48 hours before picking), L - glutamine is 0.1% - 0.2%, and the glucose is decreased to 0.08% - 0.12%.

[0039] In the above embodiments, the yeast extract can be selected as a high-purity yeast extract product. Glucose is selected as food-grade glucose. Magnesium sulfate is of analytical purity grade. Potassium dihydrogen phosphate is selected as an agricultural-grade or analytical-purity-grade product. L-arginine, L-glutamine, and zinc sulfate can also be selected as analytical-purity-grade products. All components can be purchased in the market. Prepare multiple clean containers for preparing the culture nutrient solutions at different stages. Specifically, during the pre-cracking stage: Exemplarily, prepare 1000 mL of the culture nutrient solution for the pre-cracking stage. Accurately weigh 0.8 g - 1.2 g of yeast extract (preferably weigh 1 g), 1.5 g - 2.5 g of glucose (preferably weigh 2 g), 0.2 g - 0.4 g of magnesium sulfate (preferably take 0.3 g), 0.3 g - 0.5 g of potassium dihydrogen phosphate (preferably weigh 0.4 g), and 0.0008 g - 0.0012 g of vitamin B1 (preferably take 0.001 g). Add the above raw materials into a container containing an appropriate amount of distilled water in sequence, stir evenly, and then make up the volume to 1000 mL with distilled water. Use a pH test paper or a pH meter to detect and adjust the pH value of the solution to between 6.0 - 6.5, for example, adjust it to pH = 6.3. During the cracking stage: Based on the 1000 mL of nutrient solution prepared in the pre-cracking stage (or it can be re-prepared). Accurately weigh 0.4 g - 0.6 g of potassium dihydrogen phosphate (such as weigh 0.5 g), and 0.8 g - 1.2 g of L-arginine (take 1 g). Add these two raw materials into the nutrient solution in the pre-cracking stage, stir well, and make them completely dissolve in the nutrient solution. During the stipe elongation stage: Re-prepare 1000 mL of the nutrient solution for the stipe elongation stage. Accurately weigh 2.5 g - 3.5 g of glucose (such as weigh 3 g), 0.04 g - 0.06 g of zinc sulfate (take 0.05 g), and other raw materials in the pre-cracking stage nutrient solution except glucose (1 g of yeast extract, 0.3 g of magnesium sulfate, 0.4 g of potassium dihydrogen phosphate, 0.001 g of vitamin B1). Add these raw materials into a container containing an appropriate amount of distilled water in sequence, stir evenly, make up the volume to 1000 mL, and adjust the pH value to between 6.0 - 6.5, such as pH = 6.2. During the pre-harvest stage: If based on the existing 1000 mL of the nutrient solution in the stipe elongation stage. Accurately weigh 1 g - 2 g of L-glutamine (such as weigh 1.5 g), and at the same time reduce the amount of glucose so that the glucose content is 0.8 g - 1.2 g (such as reduce to 1 g). Add L-glutamine into the nutrient solution, stir evenly, after adjusting the glucose content, detect and finely adjust the pH value to between 6.0 - 6.5 again, such as pH = 6.4. According to the physiological requirements of Dictyophora rubrovolvata at different growth stages such as the pre-cracking stage, cracking stage, stipe elongation stage, and pre-harvest stage, accurately adjust the components of the culture nutrient solution.Provide basic nutrition in the early stage of shell breaking, add specific nutritional components during the shell breaking period to promote the shell breaking process, up-regulate the glucose content and add zinc sulfate during the stipe elongation period to meet the energy and trace element requirements for stipe growth, and adjust the nutritional components in the pre-harvest period to help improve the quality and storage tolerance of Dictyophora rubrovolvata, thereby improving the overall growth quality and yield of Dictyophora rubrovolvata. Appropriate nutrient supply can accelerate the growth process of Dictyophora rubrovolvata and reduce stagnation or poor growth phenomena during the growth cycle. For example, during the stipe elongation period, sufficient and appropriate nutrients enable the stipe to elongate quickly and healthily, shortening the entire growth cycle and improving production efficiency.

[0040] In one specific embodiment, the pine needles laid on the microporous ceramic plate need to be pretreated, and the pretreatment method is as follows: A. Sterilize the pine needles in steam at 80 - 100 °C for 30 - 60 minutes; B. Immerse the sterilized pine needles in the pine needle nutrient solution for 6 - 12 hours. The components of the pine needle nutrient solution are 0.05% - 0.1% yeast extract, 0.1% - 0.3% glucose, 0.01% - 0.05% magnesium sulfate, 0.02% - 0.06% potassium dihydrogen phosphate, and the balance is water.

[0041] In the above embodiment, a conventional high-pressure steam sterilizer is selected, and a clean plastic bucket or glass container is prepared as a container for soaking pine needles. The size is selected according to the amount of pine needles to be processed, ensuring that the pine needles can be completely immersed in the nutrient solution and have a certain amount of space for movement. Yeast extract, glucose, magnesium sulfate and potassium dihydrogen phosphate can all be purchased on the market or from a specialized company. Gather an appropriate amount of fresh pine needles, remove impurities and obviously damaged parts, and arrange them into bundles or place them loosely on the rack in the high-pressure steam sterilizer. The temperature of the high-pressure steam sterilizer is set to 90°C, and the sterilization time is set to 45 minutes. Start the sterilizer, and start timing after the temperature reaches the set value and stabilizes, ensuring that the pine needles are continuously sterilized at the specified temperature for 45 minutes. After the sterilization is completed, wait until the pressure in the pot naturally drops to zero, open the lid and take out the pine needles. Prepare pine needle nutrient solution, and prepare 10L pine needle nutrient solution. Accurately weigh 7.5g, 20g glucose, 3g magnesium sulfate, and 4g potassium dihydrogen phosphate. Add these raw materials to the soaking container with appropriate amount of distilled water in turn, stir evenly, and then dilute to 10L with distilled water. Prepare pine needle nutrient solution, assuming that 10L pine needle nutrient solution is prepared. Accurately weigh 7.5g yeast extract, 20g glucose, 3g magnesium sulfate, and 4g potassium dihydrogen phosphate. Add these raw materials to the soaking container with appropriate amount of distilled water in turn, stir evenly, and then dilute to 10L with distilled water. Immerse the steam-sterilized pine needles completely in the prepared pine needle nutrient solution and soak for 9 hours. During the soaking process, stir appropriately or turn the pine needles every once in a while to ensure that the pine needles absorb the nutrient solution evenly. After the soaking is completed, remove the pine needles from the nutrient solution, drain the excess liquid, and use them to lay on the microporous ceramic plate. By sterilizing in 80-100℃ steam for 30-60 minutes, bacteria, fungi and other harmful microorganisms that may be carried on the surface and inside of the pine needles can be effectively killed. If these microorganisms are not removed, they may compete with bamboo fungus for nutrients during the cultivation of red bamboo fungus, and even cause diseases, affecting the growth and fruiting of bamboo fungus. Sterilization treatment creates a relatively sterile growth environment for red bamboo fungus. The sterilized pine needles are soaked in a pine needle nutrient solution with specific components to allow the pine needles to absorb nutrients such as yeast extract, glucose, magnesium sulfate, and potassium dihydrogen phosphate. When the pine needles treated in this way are laid on a microporous ceramic plate for the cultivation of red bamboo fungus, the pine needles can not only serve as a physical support structure, but also slowly release the absorbed nutrients, providing an additional nutrient source for the red bamboo fungus buds, helping to promote the growth and development of red bamboo fungus and improve its fruiting quality and yield.

[0042] In one specific embodiment, a 100-mesh filter is provided at the inlet end of the first pipeline; sterile air is continuously introduced into the culture nutrient solution during the infiltration stage, and the vent hole is located on the side wall of the liquid storage layer 1 cm away from the bottom plate.

[0043] In the above embodiments, the 100-mesh filter screen can be made of stainless steel, which has good corrosion resistance and can adapt to the chemical environment of the culture nutrient solution. At the inlet end of the first pipeline, the selected 100-mesh stainless steel filter screen is tightly installed through a suitable connector (such as a pipe clamp). An air filter is used to prepare sterile air. For example, a high-efficiency particulate air filter (HEPA) is selected, which can effectively filter impurities such as bacteria and dust particles in the air and ensure that the output air meets the sterile standard. At a position 1 cm from the bottom plate on the side wall of the liquid storage layer, a vent hole with an appropriate diameter (such as 0.5 cm - 1 cm) is drilled using a drilling tool. The number of vent holes is reasonably determined according to the size of the liquid storage layer and the volume of the culture nutrient solution to ensure that sterile air can be evenly and effectively introduced into the nutrient solution. Then, the sterile air delivery pipeline is connected to the vent hole to ensure good sealing and prevent leakage of the nutrient solution and entry of external air. Continuously introducing sterile air during the infiltration stage of the culture nutrient solution can increase the dissolved oxygen content in the nutrient solution. Dictyophora rubrovolvata needs to carry out respiration during its growth process, and sufficient dissolved oxygen is beneficial to the normal progress of its metabolic activities and promotes the growth and development of the mushroom buds. The vent hole is located 1 cm from the bottom plate on the side wall of the liquid storage layer, allowing sterile air to be introduced from the bottom, which can better mix with the nutrient solution, improve the dissolved oxygen efficiency, and avoid excessive agitation of the nutrient solution caused by improper ventilation position, affecting the stability of the mushroom buds.

[0044] In one specific embodiment, it further includes adjusting environmental parameters, specifically: When the decline rate of the red edge index REI reaches 8% - 15% and it is determined that the outer skin of the mushroom bud begins to rupture, the temperature in the incubator is lowered by 2 - 3 °C, and at the same time, the light intensity is adjusted to 200 - 300 lux; The air circulation rate in the incubator is increased to 0.5 - 0.8 m / s to ensure sufficient oxygen supply and promote the growth of the mushroom stalk.

[0045] In the above-described embodiments, when the decline rate of the red-edge index REI reaches a specific range, it indicates that the outer skin of the mushroom bud begins to rupture. At this time, the temperature inside the incubator is lowered by 2 - 3°C, simulating the slight temperature changes that may occur around the red-veiled stinkhorn when it breaks out of its shell in the natural environment, providing suitable temperature conditions for the subsequent growth of the mushroom bud, conforming to its growth habits, and helping the mushroom bud smoothly pass through the shell-breaking period and transition to the stage of stipe elongation. Increasing the air circulation rate inside the incubator to 0.5 - 0.8 m / s can ensure sufficient oxygen supply. During the growth process of the red-veiled stinkhorn, especially during the stipe elongation period, the respiratory function is vigorous and the demand for oxygen increases. Sufficient oxygen can accelerate its cell respiration, provide more energy for the growth of the stipe, promote the division and elongation of stipe cells, enable the stipe to grow rapidly and robustly, and thus improve the overall quality and commercial value of the red-veiled stinkhorn. By comprehensively adjusting environmental parameters such as temperature, light intensity, and air circulation rate, a microenvironment more suitable for the growth of the red-veiled stinkhorn is created, reducing growth abnormalities or stagnation caused by unsuitable environments, improving the stability and success rate of the red-veiled stinkhorn cultivation process, and facilitating large-scale and efficient cultivation.

[0046] In one specific embodiment, if the shell-breaking is delayed, the decline rate of REI < 0.5% / h lasts for 24 hours and NDWI does not increase, then: Gently scratch the top of the mushroom bud with a sterile needle tip, or slightly vibrate the surface of the mushroom bud through ultrasonic waves; Simultaneously raise the humidity of the cultivation layer to 90%.

[0047] In the above-described embodiments, gently scratching the top of the mushroom bud with a sterile needle tip can break the original structure on the surface of the mushroom bud. To a certain extent, it simulates the external stimuli that may be encountered in the natural environment and stimulates the shell-breaking mechanism of the mushroom bud. Or slightly vibrate the surface of the mushroom bud through ultrasonic waves. Utilizing the mechanical effect of ultrasonic waves, external stimuli are given to the mushroom bud from the outside, triggering physiological changes inside the mushroom bud, helping to break the stagnant state of its growth, promoting the rupture of the outer skin of the mushroom bud, and increasing the possibility of shell-breaking.

[0048] A specific example is given below.

[0049] Collect 8-mature red-veiled stinkhorn mushroom buds. These mushroom buds are in a plump oval shape, with an obvious bulge at the top and a hard texture. After testing, their moisture content is about 73%. The mushroom buds are intact as a whole and show no signs of any diseases. During harvesting, the staff carefully cut the mushroom buds from the roots with a sharp knife, and then place them in an open plastic box with pine needles spread at the bottom, ensuring that the roots of the mushroom buds face down and the tops face up. When multiple layers need to be placed, pine needles are used to separate them between the tops and roots of the mushroom buds to avoid mutual extrusion damage. After filling, the box is placed with an open mouth and not sealed.

[0050] After the collection is completed, immediately move the box containing the mushroom buds to the transport vehicle. The refrigeration equipment in the transport vehicle is turned on in advance, and the temperature is controlled at 15°C and the humidity is controlled at 75%. Such an environment can ensure the quality of the mushroom buds during transportation. After about 2 hours of transportation, the mushroom buds are successfully delivered to the cultivation laboratory.

[0051] In the cultivation laboratory, an incubator made of transparent glass material is selected. The top of this incubator is open, which is conducive to air circulation. A microporous ceramic plate with a pore diameter of 30μm and a porosity of 50% is installed on the bottom plate of the incubator. It is installed at a vertical distance of 3 cm from the bottom plate of the incubator, thereby dividing the incubator into an upper cultivation layer and a lower liquid storage layer.

[0052] The pine needles are processed in advance: The needles of local common pine trees are collected. After selecting impurities and damaged parts, the pine needles are put into a high-pressure steam sterilizer. The temperature of the sterilizer is set at 90°C and the sterilization time is set at 45 minutes. After the sterilization is completed and the pressure in the pot naturally drops to zero, take out the pine needles and soak them in the pine needle nutrient solution. The pine needle nutrient solution is prepared according to the ratio of 0.075% yeast extract, 0.2% glucose, 0.03% magnesium sulfate, and 0.04% potassium dihydrogen phosphate, and the balance is water. The pine needles are soaked in the nutrient solution for 9 hours, and they are stirred at regular intervals during this period to ensure that they can evenly absorb the nutrients. After the soaking is over, take out the pine needles and drain them. A layer of pine needle layer with a thickness of 4 cm is laid on the microporous ceramic plate at a laying density of 0.2 g / cm³.

[0053] After the pine needle laying is completed, the collected 8-mature Dictyophora rubrovolvata mushroom buds are carefully placed on the pine needles. During the entire cultivation process, the temperature of the cultivation environment is accurately maintained at 23°C with the help of the air-conditioning system. At the same time, a contact liquid level sensor is installed at a suitable position on the side wall of the liquid storage layer to monitor the height of the cultivation nutrient solution in the liquid storage layer in real time and transmit the data to the control center in real time.

[0054] The supply of the cultivation nutrient solution adopts a circulating method. A liquid inlet and an outlet made of plastic material are provided on the side wall of the liquid storage layer. The first pipeline and the second pipeline are selected as acid and alkali resistant PVC plastic hoses with a pipe diameter of 8 mm. A small peristaltic pump and a normally closed first solenoid valve are installed in the first pipeline, and a normally closed second solenoid valve is installed in the second pipeline, and the height of the liquid storage layer is higher than that of the external container. The liquid inlet is connected to an external container containing the cultivation nutrient solution through the first pipeline, and the liquid outlet is also connected to the external container through the second pipeline.

[0055] At 9:00 every morning and 3:00 in the afternoon, the control center starts the nutrient solution circulation program. The first solenoid valve and the peristaltic pump are turned on, and the second solenoid valve is turned off. The peristaltic pump pumps the culture nutrient solution in the external container into the liquid storage layer through the first pipeline. When the liquid level sensor detects that the liquid level rises to contact the lower surface of the microporous ceramic plate (the liquid level is 3 cm at this time), the control center immediately closes the first solenoid valve and the peristaltic pump to stop the liquid injection. After that, the nutrient solution slowly wets the pine needles through the capillary action of the microporous ceramic plate, and the infiltration thickness is about 1 / 2 of the pine needle thickness (i.e., 2 cm), avoiding submerging the mushroom buds. After 15 minutes, the control center opens the second solenoid valve, and the culture nutrient solution in the liquid storage layer drains back to the external container under the action of gravity, completing one cycle.

[0056] During the cultivation process, the growth of Dictyophora rubrovolvata is monitored by a Specim IQ hyperspectral imager set at the center of the top of the incubator. The imager is 40 cm away from the mushroom buds, and the field of view can cover the entire incubator. It is set that the imager collects hyperspectral images every 2 hours, and the collected images are analyzed by the supporting data analysis software after collection.

[0057] On the 1st day of cultivation, the reflectance data of the volva area is extracted from the visible light band image, and the red edge index REI is calculated. At this time, R600 is 0.35 and R700 is 0.45. According to the formula REI = (R700 - R600) / (R700 + R600), then REI = (0.45 - 0.35) / (0.45 + 0.35) = 0.125. By the 2nd day, REI is calculated again. R600 becomes 0.3 and R700 becomes 0.4. REI = (0.4 - 0.3) / (0.4 + 0.3) ≈ 0.143. During this period, the decline rate of REI is about (0.125 - 0.143) / 24 ≈ -7.5% / h, which has not reached the hatching standard. Continuing the monitoring, on the 3rd day, R600 becomes 0.25 and R700 becomes 0.35. At this time, REI = (0.35 - 0.25) / (0.35 + 0.25) ≈ 0.167. From the 2nd day to the 3rd day, the decline rate of REI is about (0.143 - 0.167) / 24 ≈ -10% / h, reaching the hatching standard.

[0058] On the 4th day, R860 was 0.4 and R1240 was 0.2. The normalized difference water index NDWI was calculated as (0.4 - 0.2) / (0.4 + 0.2) = 0.33. As the cultivation time passed, by the 5th day, R860 became 0.45 and R1240 became 0.18. Then NDWI = (0.45 - 0.18) / (0.45 + 0.18) = 0.43. The continuous increase in NDWI indicated that the stipe entered the rapid growth stage.

[0059] When the cultivation reached the 8th day, the short-wave infrared band images were analyzed. After preprocessing such as image enhancement and threshold segmentation, the area A of the skirt region was calculated as 5.5 cm², the perimeter P was 8.5 cm, and the skirt expansion index SVI = 4π×5.5 / 8.5²≈ 0.96. At the same time, the drooping angle of the skirt was calculated by the vector method. The top center point, the outermost drooping point, and the skirt support point (the connection between the stipe and the volva) of the skirt were identified, and the vector included angle was calculated, and the drooping angle of the skirt was about 135°. Since SVI > 0.3 and the drooping angle of the skirt > 120°, it indicated that Dictyophora rubrovolvata had reached the standard for collection.

[0060] During this cultivation process, the components of the cultivation nutrient solution were adjusted accordingly for different growth stages. Before the shell-breaking stage, 1000 mL of the cultivation nutrient solution was prepared. 1 g of yeast extract, 2 g of glucose, 0.3 g of magnesium sulfate, 0.4 g of potassium dihydrogen phosphate, and 0.001 g of vitamin B1 were accurately weighed, added with an appropriate amount of distilled water, stirred evenly, and then made up to 1000 mL, and the pH value was adjusted to 6.3 with a pH meter. During the shell-breaking stage, on the basis of the 1000 mL nutrient solution in the pre-shell-breaking stage, 0.5 g of potassium dihydrogen phosphate and 1 g of L-arginine were added, and stirred evenly until completely dissolved. During the stipe elongation stage, 1000 mL of the nutrient solution was freshly prepared. 3 g of glucose, 0.05 g of zinc sulfate, and other raw materials except glucose in the pre-shell-breaking stage were weighed, stirred evenly, made up to 1000 mL, and the pH value was adjusted to 6.2. Before the harvesting stage (48 hours before picking), on the basis of the 1000 mL nutrient solution in the stipe elongation stage, 1.5 g of L-glutamine was added, and at the same time the glucose content was reduced to 1 g, stirred evenly, and the pH value was slightly adjusted to 6.4.

[0061] In addition, a 100-mesh stainless steel filter screen was installed at the inlet end of the first pipeline to prevent impurities from entering the liquid storage layer. At a position 1 cm from the bottom plate on the side wall of the liquid storage layer, 4 ventilation holes with a diameter of 0.8 cm were evenly drilled. Sterile air was prepared by a high-efficiency particulate air (HEPA) filter and continuously introduced into the nutrient solution to increase the dissolved oxygen content in the nutrient solution and promote the growth of Dictyophora rubrovolvata.

[0062] In this embodiment, the eight-ripe mushroom buds collected are cultured to produce mushrooms. The mushroom yield rate is 97% within 5 to 8 days, the intact mushroom shape rate is over 94%, and the mushroom yield rate reaches 56% within 4 days. Moreover, there are no significant differences in terms of the fresh product size, fresh product quality, fresh product texture characteristics, and nutritional components of the cultivated Dictyophora rubrovolvata compared with those of the mushrooms grown in the forest.

[0063] The equipment quantities and processing scales described here are used to simplify the description of the present invention. The applications, modifications, and variations of the present invention are obvious to those skilled in the art.

[0064] Although the embodiments of the present invention have been disclosed as above, it is not limited to only the applications listed in the description and embodiments. It can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily achieved. Therefore, without departing from the general concept defined by the claims and the equivalent scope, the present invention is not limited to the specific details and the illustrated examples described herein.

Claims

1. A method for in vitro differentiation and fruiting of red bamboo fungus buds, characterized in that: include: Harvest 7-9 mature red stem bamboo fungus buds at the red stem bamboo fungus planting site; Cultivating 7-9 mature red-stemmed bamboo fungus buds, specifically: arranging a microporous ceramic plate on the bottom plate of a culture box, so that the culture box forms a culture layer located on the top and a liquid storage layer located on the bottom, laying a layer of pine needles on the microporous ceramic plate, placing the collected 7-9 mature red-stemmed bamboo fungus buds on the pine needles, and maintaining the culture environment temperature at 21-27° C.; The liquid storage layer is regularly filled with culture nutrient solution, so that the roots of the red-stemmed bamboo fungus buds and the laid pine needles are intermittently in contact with the culture nutrient solution, and nutrients and water are added to the red-stemmed bamboo fungus buds; A multispectral imager is arranged at the top center of the incubator to monitor the growth of the red-stemmed bamboo fungus at different stages through the multispectral imager, and the monitoring data is used to obtain in real time whether the buds of the red-stemmed bamboo fungus have broken, whether the stems have elongated, and whether they can be picked.

2. The method for in vitro differentiation and fruiting of red bamboo fungus buds as claimed in claim 1, characterized in that: The culture nutrient solution comprises the following components at different growth stages of Dictyophora rubra: In the early stage of shell breaking, yeast extract 0.08%-0.12%, glucose 0.15%-0.25%, magnesium sulfate 0.02%-0.04%, potassium dihydrogen phosphate 0.03%-0.05%, vitamin B1 0.0008%-0.0012%, pH 6.0-6.5, the balance is water; During the shell-breaking period, add 0.04%-0.06% potassium dihydrogen phosphate and 0.08%-0.12% L-arginine; During the stipe elongation period, glucose was increased to 0.25%-0.35%, and zinc sulfate was 0.004%-0.006%; In the early stage of harvest, L-glutamine is adjusted to 0.1%-0.2%, and glucose is reduced to 0.08%-0.12%.

3. The method for in vitro differentiation and fruiting of red bamboo fungus buds as claimed in claim 1, characterized in that: The pine needles laid on the microporous ceramic plate need to be pre-treated, and the pre-treatment method is as follows: A. Sterilize the pine needles in 80-100℃ steam for 30-60 minutes; B. Soak the sterilized pine needles in a pine needle nutrient solution for 6-12 hours. The pine needle nutrient solution comprises 0.05%-0.1% yeast extract, 0.1%-0.3% glucose, 0.01%-0.05% magnesium sulfate, 0.02%-0.06% potassium dihydrogen phosphate, and the balance is water.

4. The method for in vitro differentiation and fruiting of red bamboo fungus buds as claimed in claim 1, characterized in that: The specific implementation of regularly filling the liquid storage layer with culture nutrient solution is as follows: The vertical distance between the microporous ceramic plate and the bottom plate of the incubator is 3-5 cm, and a liquid level sensor is arranged on the side wall of the liquid storage layer; A liquid inlet and a liquid outlet are arranged on the side wall of the liquid storage layer, the liquid inlet is connected to an external container through a first pipeline, the liquid outlet is connected to the external container through a second pipeline, a micro pump and a first electromagnetic valve are arranged in the first pipeline, a second electromagnetic valve is arranged in the second pipeline, and the height of the liquid storage layer is higher than the height of the external container; Cultivation nutrient solution rising stage: open the first solenoid valve and the micro pump, close the second solenoid valve, inject the cultivation nutrient solution into the liquid storage layer, and when the liquid level sensor detects that the liquid level rises to contact the lower surface of the microporous ceramic plate, close the first solenoid valve and the micro pump, stop the liquid injection, and then slowly wet the pine needles through the capillary action of the microporous ceramic plate without immersing the buds; Cultivation nutrient solution descending stage: opening the second solenoid valve to discharge the cultivation nutrient solution of the liquid storage layer into the external container; The process is circulated 1-3 times a day, and the liquid level is maintained for 5-20 minutes each time. The height of the nutrient solution is 1 / 3-1 / 2 of the thickness of the pine needles during soaking to avoid completely submerging the buds. The incubator is an incubator with an open top.

5. The method for in vitro differentiation and fruiting of red bamboo fungus buds as claimed in claim 4, characterized in that: A 100-mesh filter is provided at the inlet end of the first pipeline; sterile air is continuously introduced into the culture nutrient solution during the infiltration stage, and the air vent is located on the side wall of the liquid storage layer 1 cm away from the bottom plate.

6. The method for in vitro differentiation and fruiting of Dictyophora rubra buds according to claim 1, characterized in that: The multispectral imager is used to monitor the growth of Dictyophora rubra at different stages, and the specific process is as follows: The multi-spectral imager is 30-50 cm away from the bacterial buds, and the field of view covers the entire incubator; The multispectral images of the bud growth process are obtained by the multispectral imager and analyzed by the analysis module, including: Obtain visible light band images to analyze the color changes of the outer skin of the mushroom buds to determine whether the shells have broken; Acquire near-infrared band images to monitor the elongation process of the stipe and determine whether the stipe is elongated; Obtain short-wave infrared band images to detect the expansion index of the fungus skirt to determine whether it can be collected.

7. The method for in vitro differentiation and fruiting of red-tuberous bamboo fungus buds as claimed in claim 6, characterized in that: The multispectral image of the fungus bud growth process is obtained by a multispectral imager and analyzed by an analysis module. The specific analysis process is as follows: The reflectance data of the trophodium area is extracted from the visible light band image, and the red edge index REI is calculated. When the red light band is 600-700nm and the REI decrease rate reaches 8%-15%, it is determined that the outer skin of the bud begins to break, that is, the shell begins to break; According to the near-infrared band images, the normalized moisture index NDWI was calculated. When the NDWI value continued to rise, it indicated that the stipe entered a rapid growth period. The fungus skirt expansion index SVI was calculated from the short-wave infrared band image. When SVI > 0.3 and the fungus skirt drooping angle > 120°, it indicated that the collection was possible.

8. The method for in vitro differentiation and fruiting of red-tuberous bamboo fungus buds as claimed in claim 7, characterized in that: It also includes adjustments to environmental parameters, specifically: When the red edge index REI decreases by 8%-15% and the outer skin of the buds begins to break, the temperature in the incubator is lowered by 2-3°C and the light intensity is adjusted to 200-300lux. Increase the air circulation rate in the incubator to 0.5 - 0.8m / s to ensure adequate oxygen supply and promote the growth of stipes.

9. The method for in vitro differentiation and fruiting of red-tuberous bamboo fungus buds as claimed in claim 8, characterized in that: If shell breakout is delayed, REI decreases at a rate of <0.5% / h for 24 hours, and NDWI does not increase, then: Use a sterile needle tip to gently scratch the top of the bud, or use ultrasound to slightly vibrate the surface of the bud; Simultaneously increase the humidity of the culture layer to 90%.

10. The method for in vitro differentiation and fruiting of Dictyophora rubra buds according to claim 1, characterized in that: The microporous ceramic plate has a pore size of 20-50 μm, a porosity of 45-55%, a pine needle laying density of 0.15-0.25 g / cm³, and a pine needle layer thickness of 3-5 cm.

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