CN113711844A

CN113711844A - Morel strain planting method

Info

Publication number: CN113711844A
Application number: CN202111180963.0A
Authority: CN
Inventors: 管燕; 廖明亮
Original assignee: Zunyi Citic Edible Fungus Professional Cooperative
Current assignee: Zunyi Citic Edible Fungus Professional Cooperative
Priority date: 2021-10-11
Filing date: 2021-10-11
Publication date: 2021-11-30
Anticipated expiration: 2041-10-11
Also published as: CN113711844B

Abstract

The invention discloses a morchella planting method, which belongs to the technical field of edible fungus cultivation and comprises the following steps: separating strains to obtain a plurality of groups of mother strains and performing differential setting; step two, producing stock seeds and cultivated seeds respectively by using the mother seeds and carrying out distinguishing setting; step three, carrying out fruiting tests on cultivated species or stock seeds, and carrying out distinguishing setting according to groups; and step four, selecting the corresponding group of mother seeds for cultivation and production according to the fruiting test result. The beneficial effect who adopts above-mentioned scheme does: the planting method of the morchella esculenta utilizes a temperature-controllable fruiting test device, can set the upper limit and the lower limit of the environmental temperature, and determines the temperature difference. Each separated test tube is marked with serial number, and the original seeds and the cultivated seeds are marked consistently by propagation. Selecting strains for production according to the fruiting time and the fruiting density of the cultivated species. Solves the problem and the effect that the morchella strains produced by growers are used for production; the strain cost for cultivating the morchella is reduced by more than one time; the stable yield and high yield of the cultivation of the morchella.

Description

Morel strain planting method

Technical Field

The invention relates to the technical field of edible mushroom cultivation, in particular to a morchella planting method.

Background

Morchella (Morchella), also known as bamboo grass, is a precious edible fungus and medicinal fungus. The upper part of the structure is in a folded net shape, which is like a honeycomb and a lamb tripe, so the name is obtained. Is a precious edible fungus and medicinal fungus. Morchella esculenta consists of a morchella-shaped cap of a gesturable cephalosome and an infertile stipe. The surface of the pileus is provided with a sub-solid layer with reticular edges, and the edges of the pileus are connected with the stipe. The stipe is cylindrical and hollow, and has smooth surface or groove.

The cap of Morchella esculenta is approximately spherical, oval to elliptical, 4-10cm high, 3-6cm wide, blunt and round at the top, and has Morchella-like pits on the surface. The pits are not shaped to be nearly circular, the width is 4-12mm, the eggshell color is light yellow brown, the ridge color is light, and the pits are irregularly crossed. The handle is approximately cylindrical, nearly white, hollow, smooth at the top, and has an enlarged base with an irregular shallow groove, 5-7cm long and about 2/3 mm thick as the pileus. Cylindrical shape of the sub-capsule, (280-320) μm. times.μ m. The spores were oblong and colorless, and 8 of them were contained in each sac and arranged in a single row. The top end of the lateral filament is enlarged and is as thick as 12 mu m.

The industrial cultivation of the morchella is characterized by high investment, high risk and high benefit. In recent years, the high benefit ratio of morchella cultivation in various regions is only below 30%, and the core reason is the technical problems of strain production technology and field cultivation technology. The manufacturing technology of the morchella is not greatly different from the production of other strains, a plurality of morchella growers can separate the strains by themselves, so that the strains are used without bad breath due to the fear of risks, only high-price strains can be purchased, and the phenomenon that the high-price morchella strains are off-spectrum is caused; even if high-price strains are purchased, the farmers have no bottom gas in mind and depend on transportation.

Disclosure of Invention

The invention aims to overcome the difficulties of the background technology and provides a morchella planting method.

In order to achieve the purpose, the technical scheme is as follows: a morchella planting method comprises the following steps: separating strains to obtain a plurality of groups of mother strains and performing differential setting; step two, producing stock seeds and cultivated seeds respectively by using the mother seeds and carrying out distinguishing setting; step three, carrying out fruiting tests on cultivated species or stock seeds, and carrying out distinguishing setting according to groups; and step four, selecting the corresponding group of mother seeds for cultivation and production according to the fruiting test result.

Further, in the first step, the separated strain is separated by polyspora, and the method comprises the following steps: step 1, preparation of a culture medium: adding 1000mL of water into 40.0-40.5 g of standard PDA culture medium, boiling, subpackaging in a test tube, a 100mL tissue culture bottle and a conical flask, keeping at 121 ℃ for 30 minutes, swinging into an inclined plane, and cooling for later use; step 2, preparing instruments and equipment: clip, blade, tweezers, blower, morchella sporophore, clean bench; and 3, performing sterile operation in a superclean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, cutting the surface of the morchella esculenta cover into small blocks with a square of 1cm by using a blade, disinfecting a clip by using alcohol to manufacture a Z-shaped hook, hooking one end of the clip on the small blocks of the morchella esculenta cover, hanging the other end of the clip on a tissue culture bottle or a conical bottle, and plugging a cotton plug; step 4, placing the tissue culture bottle or the conical flask in a constant temperature cabinet at 20 ℃ for 12-24 hours, taking out the hook and the tissue block in a superclean bench, continuously covering a plug, and culturing at 20 ℃; and 5, observing and identifying bacterial colonies germinated by the spores, selecting a small amount of hyphae to transfer into a test tube, numbering and marking, selecting 7-14 numbered and propagating marks, and inoculating 1 seed to each 7-14 numbered and marked mother seeds.

Further, in the first step, the strain isolation adopts tissue isolation, and the method comprises the following steps: step a, preparing a culture medium: adding 1000ml of water into 40.0-40.5 g of standard PDA culture medium, boiling, subpackaging in test tubes, keeping at 121 ℃ for 30 minutes, forming an inclined plane, and cooling for later use; step b, preparing instruments and equipment: blades, tweezers, a blower, morchella sporocarp, an ultra-clean workbench and an alcohol lamp; and c, performing aseptic operation in a clean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, wiping the surface of the morchella esculenta and instruments by using a 75% alcohol cotton ball, sterilizing by using an instrument alcohol lamp flame, longitudinally cutting off morchella sporophores, picking tissues at the joint of the handle and the cover, placing the tissues into a slant culture medium test tube, and numbering and marking; and d, placing the culture medium test tube in a constant-temperature incubator at 20 ℃ for culturing for 2-5 days, selecting the germination hyphae for tube transfer culture, marking according to the number, selecting 7-14 serial number propagation markers, and inoculating 1 stock seed to each 7-14 serial number marked stock seeds.

Further, in the second step, the production of the stock comprises the following steps: step A, preparation of a culture medium: soaking wheat grains in lime for 24 hr or boiling, maintaining for about 30 min, adding sawdust, soil, gypsum, etc., mixing, bottling, sterilizing and cooling; step B, inoculating and culturing: 7-14 slant mother seeds marked by the serial number of polyspora separation or tissue separation are used, 1 bottle of stock culture medium is inoculated to each mother seed for about 10 days of culture, and the mother seeds are marked according to the serial numbers.

Further, the production of the cultivar comprises the following steps: firstly, preparing a culture medium: soaking wheat grains in lime for 24 hr or boiling, maintaining for about 30 min, adding sawdust, soil, gypsum, etc., mixing, bottling, sterilizing and cooling; step two, inoculating and culturing: the stock seeds marked with the 9 bottles of numbers are inoculated into 1 cultivation bag in 1 bottle, and are cultured for about 15 days, and are marked according to the stock seed numbers.

Further, the fruiting test comprises the following steps: step (I), setting a fruiting environment: setting the upper temperature limit of 19 ℃, the lower temperature limit of 10 ℃ and the temperature difference of 9 ℃; step two, selecting a fruiting container; selecting soil and sowing: and (3) disinfecting soil by using lime, putting the disinfected soil into a cultivation frame, humidifying, broadcasting cultivated species, marking according to the serial number of the cultivated species, sowing 7-14 frames according to the marked serial number of strains, and screening 7-14 cultivated species. (ii) a Step four, sowing for about 5 days, and placing a nutrition bag; step five, humidity management is carried out for about 30 days; and step six, analyzing the fruiting test condition of the morchella.

Further, the manufacturing method of the nutrition bag comprises the steps of soaking wheat grains in lime water for 12-24 hours, adding water into corncobs, turning over once for 24 hours, sealing for 48 hours, uniformly mixing with lime, gypsum and the soaked wheat grains, filling into a 12 x 24 polypropylene bag, sterilizing at high temperature and high pressure for 8 hours, and cooling for later use; each bag of the nutrition bag has the wet weight of about 350 g.

Further, the nutrition bag has the following formula: 96-104 g of corncobs, 146-154 g of wheat grains, 1-3 g of gypsum and 1-3 g of lime.

Furthermore, the fruiting test device of the morchella planting method comprises a plurality of temperature adjusting devices, a plurality of humidity adjusting devices, a fruiting container, a seed spreading device and a nutrient solution container, wherein the fruiting containers are arranged at the bottom of the test room and comprise a container body and a cover body, and the bottom of the container body is provided with a water draining hole; the cover body is of a double-layer structure, a rotating shaft is arranged at the center of the cover body, a fan-shaped window is arranged on the double-layer structure, and the size of the window can be adjusted by rotating the double-layer structure of the cover body relatively.

The beneficial effect who adopts above-mentioned scheme does: the planting method of the morchella esculenta utilizes a temperature-controllable fruiting test device, can set the upper limit and the lower limit of the environmental temperature, and determines the temperature difference. Each separated test tube is marked with serial number, and the original seeds and the cultivated seeds are marked consistently by propagation. Selecting strains for production according to the fruiting time and the fruiting density of the cultivated species. Solves the problem and the effect that the morchella strains produced by growers are used for production; the strain cost for cultivating the morchella is reduced by more than one time; the stable yield and high yield of the cultivation of the morchella.

Drawings

Fig. 1 is a schematic structural view of a fruiting container in a front cross section of the morchella planting method of the present invention.

Fig. 2 is a schematic top view of a fruiting container of the morchella planting method of the present invention.

Fig. 3 is a schematic view of a three-dimensional structure of a fruiting container of the morchella planting method of the present invention.

Fig. 4 is a schematic view of a three-dimensional structure of a fruiting test device of the morchella planting method of the present invention.

In the figure, 1-a body, 2-an upper cover body, 3-a lower cover body, 4-a draining plate, 5-a window cover, 6-a window, 7-a rotating shaft, 8-soil, 9-a temperature adjusting device, 10-a humidity adjusting device, 11-a fruiting container, 12-a nutrient solution container and 13-a seed scattering device.

Detailed Description

The technical solution of the present invention will be clearly and completely described below with reference to the specific embodiments of the present invention. The described embodiments are only some, not all embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The first embodiment is as follows:

a morchella planting method comprises the following steps: separating strains to obtain a plurality of groups of mother strains and performing differential setting; step two, producing stock seeds and cultivated seeds respectively by using the mother seeds and carrying out distinguishing setting; step three, carrying out fruiting tests on cultivated species or stock seeds, and carrying out distinguishing setting according to groups; and step four, selecting the corresponding group of mother seeds for cultivation and production according to the fruiting test result.

The industrial cultivation of the morchella is high in investment, high in risk and high in benefit, and in recent years, the proportion of the high benefit of the morchella cultivation in various regions is only below 30%. The manufacturing technology of the morchella is not greatly different from the production of other strains, a plurality of morchella growers can separate the strains by themselves, so that the strains are used without bad breath due to the fear of risks, only high-price strains can be purchased, and the phenomenon that the high-price morchella strains are off-spectrum is caused; even if high-price strains are purchased, the heart is free of bottom gas and depends on transportation.

The planting method of the morchella esculenta utilizes a temperature-controllable fruiting test device, can set the upper limit and the lower limit of the environmental temperature, and determines the temperature difference. Each separated test tube is marked with serial number, and the original seeds and the cultivated seeds are marked consistently by propagation. Selecting strains for production according to the fruiting time and the fruiting density of the cultivated species. Solves the problem and the effect that the morchella strains produced by growers are used for production; the strain cost for cultivating the morchella is reduced by more than one time; the stable yield and high yield of the cultivation of the morchella.

Example two:

Preferably, in the first step, the separated strain is separated by polyspora, and the method comprises the following steps: step 1, preparation of a culture medium: adding water 1000mL into standard PDA culture medium 40.0g, boiling, subpackaging in test tube and 100mL tissue culture bottle, conical flask, maintaining at 121 deg.C for 30 min, placing into inclined plane, and cooling; step 2, preparing instruments and equipment: clip, blade, tweezers, blower, morchella sporophore, clean bench; and 3, performing sterile operation in a superclean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, cutting the surface of the morchella esculenta cover into small blocks with a square of 1cm by using a blade, disinfecting a clip by using alcohol to manufacture a Z-shaped hook, hooking one end of the clip on the small blocks of the morchella esculenta cover, hanging the other end of the clip on a tissue culture bottle or a conical bottle, and plugging a cotton plug; step 4, placing the tissue culture bottle or the conical flask in a constant temperature cabinet at 20 ℃ for 12-24 hours, taking out the hook and the tissue block in a superclean bench, continuously covering a plug, and culturing at 20 ℃; and 5, observing and recognizing bacterial colonies germinated from the spores, selecting a small amount of hyphae to transfer into a test tube, numbering and marking, selecting 7 numbered and expanding propagation marks, and inoculating 1 seed to each 7 numbered and marked mother seeds.

The spore separation method in the mother strain separation method is a method for obtaining a bacterial strain by germinating sexual spores or asexual spores of edible fungi into hyphae. The separation method is divided into a single spore separation method and a multi-spore separation method according to the different numbers of picked spores during separation. Some edible fungi have no sexual difference of spores, and pure strains are obtained by adopting monospore separation and have fructification capability; and other edible fungi have different spores, so hyphae obtained by separating monospores cannot fruit. The production adopts a polysporus separation method, and a plurality of spores are inoculated on the same culture medium, germinated and naturally mated to obtain pure seeds.

The mother strain is the key of strain production, the required purity is high, mixed bacteria cannot be generated, and meanwhile, the mycelium of the mother strain is fine and weak in capability of decomposing compost. Therefore, the mother strain mycelium needs to be cultured on an agar culture medium which is rich in nutrition, easy to absorb and smooth in surface, and easy to identify whether the mixed bacteria infection exists. PDA culture Medium is a short for Potato glucose Agar culture Medium, namely, Potato Dextrose Agar (Medium), which corresponds to Potato, glucose and Agar in English in sequence. In the scheme of the invention, through the test and comparison of the inventor, 40.0-40.3 (g) of the dosage of the PDA culture medium is selected, and the culture success rate of the mother seeds is shown in table 1.

Table 1: comparison table of PDA culture medium dosage and culture success rate in morel planting method

Dosage of PDA culture medium (g)	＜40.0	40.0	40.1	40.3	＞40.3
						Culture success rate (%)	＜90	95	98	96	＜92

The selection of mother seed separating material, whether used as sporophore for spore separation or tissue separation, should select large and robust single-grown sporophore as separating material from population with high yield, good growth vigor, strong adaptability and no mixed bacteria and insect pest. The excellent strain mother seeds obtained by separation need to be propagated and cultured in an enlarged way because the quantity is limited and the production requirement cannot be met. The method comprises cutting slant culture medium and mycelium into small pieces with short rice grain size, transferring onto new slant culture medium, culturing at proper temperature, and allowing mycelium to grow on the slant. According to the experimental method, the inventor tests and analyses show that the single successful sampling probability of the sporophore separation mother seeds selected as the mother seed separation material is 28-29%. The probability of success at least once in 7 samples is about 90%; the probability of 14 samples being successful at least once is about 99%. The probability of 9 samples being successful at least once is slightly higher than 95%. For a specific comparison see table 2.

Table 2: comparison table of mother seed separation propagation test tube count and pure seed obtaining success rate in morchella planting method

Propagation test tube count (only)	＜7	7	9	14	＞14
						Success rate of obtaining pure breed (%)	＜90	90	95	99	＞99

Preferably, in the second step, the production of the stock comprises the following steps: step A, preparation of a culture medium: soaking wheat grains in lime for 24 hr or boiling for about 30 min to make wheat grains absorb water and wheat bran not crack, adding sawdust (0.5 CM), soil, gypsum, etc., mixing, bottling, sterilizing, and cooling; the formula of the culture medium is as follows: 68% of wood chips, 22% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step B, inoculating and culturing: inoculating 1 bottle of stock culture medium to each 7 slant mother strains with serial numbers of multi-spore separation for culture, and marking according to the serial numbers of the mother strains to obtain 7 bottles in total; the strain bottle is full of strains after about 10 days.

Preferably, in the second step, the production of cultivars comprises the following steps: firstly, preparing a culture medium: soaking wheat grains in lime for 24 hr or boiling, maintaining for about 30 min, adding sawdust, soil, gypsum, etc., mixing, bottling, sterilizing and cooling; the formula of the culture medium is as follows: 45% of wood chips, 45% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step two, inoculating and culturing: inoculating 1 bottle of stock seeds marked by 7 bottles of numbers into 1 cultivation bag, culturing, marking according to the stock seeds numbers, and totally packaging 7 bags; the strain bags are full of the strain bags in about 15 days.

Example three:

Preferably, in the first step, the separated strain is separated by polyspora, and the method comprises the following steps: step 1, preparation of a culture medium: adding water 1000mL into standard PDA culture medium 40.3g, boiling, subpackaging in test tube and 100mL tissue culture bottle, conical flask, maintaining at 121 deg.C for 30 min, placing into inclined plane, and cooling; step 2, preparing instruments and equipment: clip, blade, tweezers, blower, morchella sporophore, clean bench; and 3, performing sterile operation in a superclean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, cutting the surface of the morchella esculenta cover into small blocks with a square of 1cm by using a blade, disinfecting a clip by using alcohol to manufacture a Z-shaped hook, hooking one end of the clip on the small blocks of the morchella esculenta cover, hanging the other end of the clip on a tissue culture bottle or a conical bottle, and plugging a cotton plug; step 4, placing the tissue culture bottle or the conical flask in a constant temperature cabinet at 20 ℃ for 12-24 hours, taking out the hook and the tissue block in a superclean bench, continuously covering a plug, and culturing at 20 ℃; and 5, observing and identifying colonies germinated by the spores, selecting a small amount of hyphae to transfer into a test tube, numbering and marking, selecting 14 numbered and expanding propagation marks, and inoculating 1 seed to each 14 numbered and marked mother seeds. The inventor tests and analyses that the probability of successful seed separation for 14 times of sampling is about 99 percent when the seed is selected as the seed separating material. However, too many sampling times will increase the workload and increase the labor cost.

Preferably, in the second step, the production of the stock comprises the following steps: step A, preparation of a culture medium: soaking wheat grains in lime for 24 hr or boiling for about 30 min to make wheat grains absorb water and wheat bran not crack, adding sawdust (0.5 CM), soil, gypsum, etc., mixing, bottling, sterilizing, and cooling; the formula of the culture medium is as follows: 68% of wood chips, 22% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step B, inoculating and culturing: inoculating 14 slant mother strains marked by the serial number of multi-spore separation into 1 bottle of stock culture medium for culture, and marking according to the serial number of the mother strains for 14 bottles in total; the strain bottle is full of strains after about 10 days.

Preferably, in the second step, the production of cultivars comprises the following steps: firstly, preparing a culture medium: soaking wheat grains in lime for 24 hr or boiling, maintaining for about 30 min, adding sawdust, soil, gypsum, etc., mixing, bottling, sterilizing and cooling; the formula of the culture medium is as follows: 45% of wood chips, 45% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step two, inoculating and culturing: inoculating 1 bottle of stock seeds marked with the number of 14 bottles into 1 cultivation bag, culturing, marking according to the number of the stock seeds, and totally packaging 14 bags; the strain bags are full of the strain bags in about 15 days.

Example four:

Preferably, in the first step, the separated strain is separated by polyspora, and the method comprises the following steps: step 1, preparation of a culture medium: adding water 1000mL into standard PDA culture medium 40.1g, boiling, subpackaging in test tube and 100mL tissue culture bottle, conical flask, maintaining at 121 deg.C for 30 min, placing into inclined plane, and cooling; step 2, preparing instruments and equipment: clip, blade, tweezers, blower, morchella sporophore, clean bench; and 3, performing sterile operation in a superclean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, cutting the surface of the morchella esculenta cover into small blocks with a square of 1cm by using a blade, disinfecting a clip by using alcohol to manufacture a Z-shaped hook, hooking one end of the clip on the small blocks of the morchella esculenta cover, hanging the other end of the clip on a tissue culture bottle or a conical bottle, and plugging a cotton plug; step 4, placing the tissue culture bottle or the conical flask in a constant temperature cabinet at 20 ℃ for 12-24 hours, taking out the hook and the tissue block in a superclean bench, continuously covering a plug, and culturing at 20 ℃; and 5, observing and recognizing bacterial colonies germinated from the spores, selecting a small amount of hyphae to transfer into a test tube, numbering and marking, selecting 9 numbered and expanding propagation marks, and inoculating 1 seed to each of the 9 numbered and marked mother seeds. The inventor tests and analyses that the probability of successful seed separation for the fruiting body selected as the seed separation material is about 95% after 9 times of sampling. Compared with 9 times of sampling, the workload, the labor cost and the success probability are balanced, and the method belongs to the optimal scheme.

Preferably, in the second step, the production of the stock comprises the following steps: step A, preparation of a culture medium: soaking wheat grains in lime for 24 hr or boiling, maintaining for about 30 min, adding sawdust, soil, gypsum, etc., mixing, bottling, sterilizing and cooling; the formula of the culture medium is as follows: 68% of wood chips, 22% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step B, inoculating and culturing: inoculating 1 bottle of stock culture medium to each 9 slant mother strains marked by the serial number of multi-spore separation for culture, and marking according to the serial number of the mother strains for 9 bottles in total; the strain bottle is full of strains after about 10 days.

Preferably, in the second step, the production of cultivars comprises the following steps: firstly, preparing a culture medium: soaking wheat grains in lime for 24 hr or boiling, maintaining for about 30 min, adding sawdust, soil, gypsum, etc., mixing, bottling, sterilizing and cooling; the formula of the culture medium is as follows: 45% of wood chips, 45% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step two, inoculating and culturing: inoculating 1 bottle of the stock seeds marked by the serial number of 9 bottles into 1 cultivation bag, culturing, marking according to the serial number of the stock seeds, and totally 9 bags; the strain bags are full of the strain bags in about 15 days.

Example five:

Preferably, in the first step, tissue isolation is adopted to isolate strains, and the method comprises the following steps: step a, preparing a culture medium: adding 1000ml of water into 40.0g of standard PDA culture medium, boiling, subpackaging in test tubes, keeping at 121 ℃ for 30 minutes, placing on an inclined plane, and cooling for later use; step b, preparing instruments and equipment: blades, tweezers, a blower, morchella sporocarp, an ultra-clean workbench and an alcohol lamp; and c, performing aseptic operation in a clean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, wiping the surface of the morchella esculenta and instruments by using a 75% alcohol cotton ball, sterilizing by using an instrument alcohol lamp flame, longitudinally cutting off morchella sporophores, picking tissues at the joint of the handle and the cover, placing the tissues into a slant culture medium test tube, and numbering and marking; and d, placing the culture medium test tube in a constant-temperature incubator at 20 ℃ for culturing for 2-5 days, selecting the germination hyphae for tube transfer culture, marking according to the number, selecting 7 numbered stock seeds for serial number propagation expanding marking, and inoculating 1 stock seed to each stock seed marked by 7 numbers.

Preferably, in the second step, the production of the stock comprises the following steps: step A, preparation of a culture medium: soaking wheat grains in lime for 24 hr or boiling for about 30 min to make wheat grains absorb water and wheat bran not crack, adding sawdust (0.5 CM), soil, gypsum, etc., mixing, bottling, sterilizing, and cooling; the formula of the culture medium is as follows: 68% of wood chips, 22% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step B, inoculating and culturing: inoculating 1 bottle of stock culture medium to each 7 slant mother seeds marked by serial numbers of tissue separation for culture, and marking according to the serial numbers of the mother seeds for 7 bottles in total; the strain bottle is full of strains after about 10 days.

The tissue separation method is a method for directly separating and culturing young tissue blocks of edible fungi into pure strains. After tissue isolation, the tubes were placed outside in an incubator for culture. And (4) after hyphae germinate around the tissue blocks and spread to the culture medium, selecting the strong hyphae for tube rotating culture. The tissue separation method is simple and convenient to operate, and is not easy to bring in mixed bacteria, and pure strains are easy to obtain.

Example six:

Preferably, in the first step, tissue isolation is adopted to isolate strains, and the method comprises the following steps: step a, preparing a culture medium: adding 1000ml of water into 40.3g of standard PDA culture medium, boiling, subpackaging in test tubes, keeping at 121 ℃ for 30 minutes, placing on an inclined plane, and cooling for later use; step b, preparing instruments and equipment: blades, tweezers, a blower, morchella sporocarp, an ultra-clean workbench and an alcohol lamp; and c, performing aseptic operation in a clean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, wiping the surface of the morchella esculenta and instruments by using a 75% alcohol cotton ball, sterilizing by using an instrument alcohol lamp flame, longitudinally cutting off morchella sporophores, picking tissues at the joint of the handle and the cover, placing the tissues into a slant culture medium test tube, and numbering and marking; and d, placing the culture medium test tube in a constant-temperature incubator at 20 ℃ for culturing for 2-5 days, selecting the germination hyphae for tube transfer culture, marking according to the number, selecting 14 serial number propagation expanding marks, and inoculating 1 seed to each 14 serial number marked mother seeds.

Preferably, in the second step, the production of the stock comprises the following steps: step A, preparation of a culture medium: soaking wheat grains in lime for 24 hr or boiling for about 30 min to make wheat grains absorb water and wheat bran not crack, adding sawdust (0.5 CM), soil, gypsum, etc., mixing, bottling, sterilizing, and cooling; the formula of the culture medium is as follows: 68% of wood chips, 22% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step B, inoculating and culturing: inoculating 14 slant mother strains marked by the serial number of tissue separation into 1 bottle of stock culture medium for culture, and marking according to the serial number of the mother strains for 14 bottles in total; the strain bottle is full of strains after about 10 days.

Example seven:

Preferably, in the first step, tissue isolation is adopted to isolate strains, and the method comprises the following steps: step a, preparing a culture medium: adding 1000ml of water into 40.1g of standard PDA culture medium, boiling, subpackaging in test tubes, keeping at 121 ℃ for 30 minutes, placing on an inclined plane, and cooling for later use; step b, preparing instruments and equipment: blades, tweezers, a blower, morchella sporocarp, an ultra-clean workbench and an alcohol lamp; and c, performing aseptic operation in a clean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, wiping the surface of the morchella esculenta and instruments by using a 75% alcohol cotton ball, sterilizing by using an instrument alcohol lamp flame, longitudinally cutting off morchella sporophores, picking tissues at the joint of the handle and the cover, placing the tissues into a slant culture medium test tube, and numbering and marking; and d, placing the culture medium test tube in a constant-temperature incubator at 20 ℃ for culturing for 2-5 days, selecting the germination hyphae for tube transfer culture, marking according to the number, selecting 9 serial number propagation expanding marks, and inoculating 1 mother seed with each of the 9 serial number marks to the original seed.

Preferably, in the second step, the production of the stock comprises the following steps: step A, preparation of a culture medium: soaking wheat grains in lime for 24 hr or boiling for about 30 min to make wheat grains absorb water and wheat bran not crack, adding sawdust (0.5 CM), soil, gypsum, etc., mixing, bottling, sterilizing, and cooling; the formula of the culture medium is as follows: 68% of wood chips, 22% of wheat grains, 7% of soil, 2% of lime and 1% of gypsum. Step B, inoculating and culturing: inoculating 1 bottle of stock culture medium to each of 9 slant mother seeds marked by serial numbers of tissue separation for culture, and marking according to the serial numbers of the mother seeds for 9 bottles in total; the strain bottle is full of strains after about 10 days.

Example eight:

according to the embodiment, the morchella planting method comprises the following steps: step (I), setting a fruiting environment: setting the upper temperature limit of 19 ℃, the lower temperature limit of 10 ℃ and the temperature difference of 9 ℃; selecting a fruiting container, and selecting various containers such as a rectangle and a square to ensure that the bottom can drain; selecting soil and sowing: disinfecting the soil with lime; the soil humidity is adjusted, the soil can be held tightly to form a cluster, and the free fall can be well dispersed; placing the sterilized soil into a cultivation frame, broadcasting cultivated species, and marking according to the number of the cultivated species; the strains were sown in 7 boxes according to the marked numbers and screened from 7 cultivars. Step four, sowing for about 5 days, and placing a nutrition bag after hyphae grow over the soil surface; and step five, covering the perforated black mulching film, and preserving heat and moisture. Checking soil humidity once a week, spraying fruiting water for about 30 days, wherein the soil water content is 50-60%; the soil bloom subsides and the primordia begin to differentiate. And step six, eliminating primordium-free or few primordiums according to test conditions such as fruiting time and fruiting density of morchella, and determining strains for production and use. A part of the selected production seeds can be propagated for production and cultivation. One part is used for preservation and cultivation in the next year.

Preferably, the nutrition bag is made of the following materials: wheat grains, corncobs, lime and gypsum. The method comprises soaking wheat grains in lime water for 12-24 hr, adding water into corn cob, turning over once for 24 hr, sealing for 48 hr, mixing with lime, gypsum and soaked wheat grains, packaging into 12 x 24 polypropylene bag, sterilizing at high temperature and high pressure for 8 hr, and cooling; each bag of the nutrition bag has the wet weight of about 350 g.

Preferably, the nutrition bag formula is as follows: 96g of corncob, 146g of wheat grain, 1g of gypsum and 1g of lime.

Artificially planted morchella needs to be supplemented by placing an external aid nutrition bag, and the main function is to provide sufficient nutrition for morchella hypha in soil, so as to be beneficial to forming more round bases and lay a foundation for the high yield of the morchella.

And 5 days or so after the morchella is sown, when the soil surface is seen to be full of hypha, the nutrition bag can be placed. Four rows of small holes are required to be drilled at the bottom of the nutrition bag, and the size of the hole is based on the standard that the nutrition materials in the nutrition bag cannot fall out. When the nutrient is placed, the hole is downward and is close to the soil surface, so that the hyphae of the morchella can slowly grow into the nutrient bag to absorb nutrition and can be transmitted to the hyphae in the soil layer. The formula value and the fruiting percentage of the nutrition bag are shown in the table 3 after the inventor tests and comparisons.

Table 3: formula value and fruiting percentage of fruiting test nutrition bag for morchella planting method

Corncob (g)	＜96	96	100	104	＞104
						Wheat (g)	＜146	146	150	154	＞154
Gypsum (g)	＜1	1	2	3	＞3
						Lime (g)	＜1	1	2	3	＞3
Percentage of fruiting (%)	＜90	91	98	93	＜91

Example nine:

according to the embodiment, the morchella planting method comprises the following steps: step (I), setting a fruiting environment: setting an upper temperature limit of 17-21 ℃, a lower temperature limit of 8-12 ℃ and a temperature difference of 7-11 ℃; step two, selecting a fruiting container; selecting soil and sowing: disinfecting soil with lime, placing the disinfected soil into a cultivation frame, humidifying, broadcasting cultivated species, marking according to the number of the cultivated species, sowing 14 frames according to the marked number of strains, and screening 14 cultivated species. (ii) a Step four, sowing for about 5 days, and placing a nutrition bag; step five, humidity management is carried out for about 30 days; and step six, analyzing the fruiting test condition of the morchella.

Preferably, the manufacturing method of the nutrition bag comprises the steps of soaking wheat grains in lime water for 12-24 hours, adding water into corncobs, turning over once for 24 hours, stewing for 48 hours, uniformly mixing with lime, gypsum and the soaked wheat grains, filling into a 12 x 24 polypropylene bag, sterilizing at high temperature and high pressure for 8 hours, and cooling for later use; each bag of the nutrition bag has the wet weight of about 350 g.

Preferably, the nutrition bag formula is as follows: 104g of corncob, 154g of wheat grain, 3g of gypsum and 3g of lime.

Example ten:

according to the embodiment, the morchella planting method comprises the following steps: step (I), setting a fruiting environment: setting an upper temperature limit of 17-21 ℃, a lower temperature limit of 8-12 ℃ and a temperature difference of 7-11 ℃; step two, selecting a fruiting container; selecting soil and sowing: disinfecting soil with lime, placing the disinfected soil into a cultivation frame, humidifying, broadcasting cultivated species, marking according to the number of the cultivated species, sowing 9 frames according to the marked number of strains, and screening from 9 cultivated species. (ii) a Step four, sowing for about 5 days, and placing a nutrition bag; step five, humidity management is carried out for about 30 days; and step six, analyzing the fruiting test condition of the morchella.

Preferably, the nutrition bag formula is as follows: 100g of corncob, 150g of wheat grain, 2g of gypsum and 2g of lime.

The technical scheme of the embodiment eight to ten has the following innovation points: setting upper and lower limits of environmental temperature by using a temperature-controllable cold storage, and determining temperature difference; each separated test tube is marked with serial number, and the original seeds and the cultivated seeds are propagated to be consistent in mark; selecting strains for production according to the fruiting time and the fruiting density of the cultivated species. The beneficial effects are that: the yield of the strain selected by the method is stable, the strain is continuously cultivated in different areas for three years, the average yield reaches more than 150kg per mu, and the average yield reaches 300kg a little; by adopting the method, the preparation of the morchella is simple and feasible, and the strain cost is reduced by more than one time; the fruiting test time of the morchella strain is controlled within 40 days.

Example eleven:

the fruiting test device 8 of the morchella planting method according to the technical scheme of the eight to ten embodiments comprises a temperature adjusting device 9, a humidity adjusting device 10, a fruiting container 11, a seed spreading device 13 and a nutrient solution container 12, wherein the fruiting containers 11 are multiple, the fruiting containers 11 are arranged at the bottom of the test device, the fruiting containers 11 comprise a container body 1 and a cover body, and the bottom of the container body 1 is provided with a draining plate 4 with draining holes; the upper part of the draining plate 4 of the container body 1 is paved with cultivation soil 7; the cover body is of a double-layer structure and comprises an upper cover body 2 and a lower cover body 3, and a rotating shaft 7 is arranged in the center; the double-layer structure is provided with a window, and the window comprises a window 6 arranged on the lower cover body and a window cover 5 arranged on the upper cover body; the size of the window can be adjusted by rotating the double-layer structure relatively. Furthermore, the window arranged on the double-layer structure is fan-shaped, the size of the fan-shaped window can be adjusted by the relative rotation of the double-layer structure of the cover body, and the size adjustment range of the window is 0-0.015 square meters. The cross-sectional area of the container body 1 is 0.21-0.25 square meter, and the height of the container body 1 is 12-18 cm.

In an eleventh embodiment, the temperature adjusting device and the humidity adjusting device arranged in the fruiting test device are symmetrically arranged along the longitudinal axis and the transverse axis of the plane of the test device respectively, and the fruiting containers are also symmetrically and uniformly distributed around the test device. The arrangement is favorable for ensuring that each fruiting container can obtain the same test environmental conditions in the test device, thereby being convenient for preferably selecting excellent strains under equal conditions.

The double-layer fan-shaped window can adjust the illumination intensity in the fruiting container within the range of completely closing to 0.015 square meter according to the progress of a fruiting test. The size of the fruiting container can be selected according to the experimental scale. The draining board can be arranged in a detachable mode so as to be convenient for cleaning and disinfecting treatment.

The seed spreading device can be set as a manual operation or mechanical automatic control device, and the volume of the nutrient solution container can be adjusted according to the size of the fruiting container. The test device can be made of a light-transmitting material.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.

Claims

1. The morchella planting method is characterized by comprising the following steps: separating strains to obtain a plurality of groups of mother strains and performing differential setting; step two, producing stock seeds and cultivated seeds respectively by using the mother seeds and carrying out distinguishing setting; step three, carrying out fruiting tests on cultivated species or stock seeds, and carrying out distinguishing setting according to groups; and step four, selecting the corresponding group of mother seeds for cultivation and production according to the fruiting test result.

2. The morchella planting method according to claim 1, wherein the method comprises the following steps: in the first step, the separated strains are separated by adopting polyspora, and the method comprises the following steps: step 1, preparation of a culture medium: adding 1000mL of water into 40.0-40.5 g of standard PDA culture medium, boiling, subpackaging in a test tube, a 100mL tissue culture bottle and a conical flask, keeping at 121 ℃ for 30 minutes, swinging into an inclined plane, and cooling for later use; step 2, preparing instruments and equipment: clip, blade, tweezers, blower, morchella sporophore, clean bench; and 3, performing sterile operation in a superclean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, cutting the surface of the morchella esculenta cover into small blocks with a square of 1cm by using a blade, disinfecting a clip by using alcohol to manufacture a Z-shaped hook, hooking one end of the clip on the small blocks of the morchella esculenta cover, hanging the other end of the clip on a tissue culture bottle or a conical bottle, and plugging a cotton plug; step 4, placing the tissue culture bottle or the conical flask in a constant temperature cabinet at 20 ℃ for 12-24 hours, taking out the hook and the tissue block in a superclean bench, continuously covering a plug, and culturing at 20 ℃; and 5, observing and identifying bacterial colonies germinated by the spores, selecting a small amount of hyphae to transfer into a test tube, numbering and marking, selecting 7-14 numbered and propagating marks, and inoculating 1 seed to each 7-14 numbered and marked mother seeds.

3. The morchella planting method according to claim 1, wherein the method comprises the following steps: in the first step, the strain separation adopts tissue separation, and comprises the following steps: step a, preparing a culture medium: adding 1000ml of water into 40.0-40.5 g of standard PDA culture medium, boiling, subpackaging in test tubes, keeping at 121 ℃ for 30 minutes, forming an inclined plane, and cooling for later use; step b, preparing instruments and equipment: blades, tweezers, a blower, morchella sporocarp, an ultra-clean workbench and an alcohol lamp; and c, performing aseptic operation in a clean bench: blowing cold air to the surface of the morchella esculenta cover by using a blower, wiping the surface of the morchella esculenta and instruments by using a 75% alcohol cotton ball, sterilizing by using an instrument alcohol lamp flame, longitudinally cutting off morchella sporophores, picking tissues at the joint of the handle and the cover, placing the tissues into a slant culture medium test tube, and numbering and marking; and d, placing the culture medium test tube in a constant-temperature incubator at 20 ℃ for culturing for 2-5 days, selecting the germination hyphae for tube transfer culture, marking according to the number, selecting 7-14 serial number propagation markers, and inoculating 1 stock seed to each 7-14 serial number marked stock seeds.

4. The morchella planting method according to claim 1, wherein the method comprises the following steps: in the second step, the production of the stock seeds comprises the following steps: step A, preparation of a culture medium: soaking wheat grains in lime for 24 hr or boiling, maintaining for about 30 min, adding sawdust, soil, gypsum, etc., mixing, bottling, sterilizing and cooling; step B, inoculating and culturing: 7-14 slant mother seeds marked by the serial number of polyspora separation or tissue separation are used, 1 bottle of stock culture medium is inoculated to each mother seed for about 10 days of culture, and the mother seeds are marked according to the serial numbers.

5. The morchella planting method according to claim 1, wherein the method comprises the following steps: in the second step, the production of cultivars comprises the following steps: firstly, preparing a culture medium: soaking wheat grains in lime for 24 hr or boiling, maintaining for about 30 min, adding sawdust, soil, gypsum, etc., mixing, bottling, sterilizing and cooling; step two, inoculating and culturing: and (3) inoculating 1 bottle of the stock seeds marked with the serial numbers of 7-14 bottles into 1 cultivation bag, culturing for about 15 days, and marking according to the serial numbers of the stock seeds.

6. The morchella planting method according to claim 1, wherein the method comprises the following steps: the fruiting test comprises the following steps: step (I), setting a fruiting environment: setting the upper temperature limit of 19 ℃, the lower temperature limit of 10 ℃ and the temperature difference of 9 ℃; step two, selecting a fruiting container; selecting soil and sowing: and (3) disinfecting soil by using lime, putting the disinfected soil into a cultivation frame, humidifying, broadcasting cultivated species, marking according to the serial number of the cultivated species, sowing 7-14 frames according to the marked serial number of strains, and screening 7-14 cultivated species. (ii) a Step four, sowing for about 5 days, and placing a nutrition bag; step five, humidity management is carried out for about 30 days; and step six, analyzing the fruiting test condition of the morchella.

7. The morchella planting method according to claim 6, wherein the method comprises the following steps: the manufacturing method of the nutrition bag comprises the steps of soaking wheat grains in lime water for 12-24 hours, adding water into corncobs, turning over once for 24 hours, stewing for 48 hours, uniformly mixing with lime, gypsum and the soaked wheat grains, filling into a 12 x 24 polypropylene bag, sterilizing at high temperature and high pressure for 8 hours, and cooling for later use; each bag of the nutrition bag has the wet weight of about 350 g.

8. The morchella planting method according to claim 6, wherein the method comprises the following steps: the formula of the nutrition bag is as follows: 96-104 g of corncobs, 146-154 g of wheat grains, 1-3 g of gypsum and 1-3 g of lime.

9. The fruiting test device of morel planting method as claimed in claim 6, wherein: comprises a temperature adjusting device, a humidity adjusting device, a fruiting container, a seed spreading device and a nutrient solution container; the mushroom growing container is arranged at the bottom of the experimental device and comprises a container body and a cover body, and a plurality of water draining holes are formed in the bottom of the container body; the cover body is of a double-layer structure, a rotating shaft is arranged at the center of the cover body, a fan-shaped window is arranged on the double-layer structure, and the size of the window can be adjusted by rotating the double-layer structure of the cover body relatively.