Excellent morchella strain as well as preparation method and application thereof
The spores of Liumei Morel were treated by normal pressure room temperature plasma (ARTP) mutagenesis technology, and the high-yield morel mutant strain M6SA24 was screened, which solved the problems of poor genetic stability and environmental adaptability of morel strains, and achieved a technological breakthrough in efficient transplantation.
Patent Information
- Application Number
- CN202510816276.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-18
- Publication Date
- 2025-08-22
AI Technical Summary
The existing morel species have poor genetic stability and weak environmental adaptability, which makes it difficult to reproduce efficient planting and the existing breeding technology has failed to make effective breakthroughs.
The spores of Liumei Morel were treated by normal pressure room temperature plasma (ARTP) mutagenesis technology. Combined with biological characteristics analysis and phenotypic screening, a high-yield morel mutant strain M6SA24 was obtained, and excellent bacterial strains were prepared.
It has improved the mycelium growth rate, early sclerosis and slow strain degeneration of morels, provided high-yield excellent strains of morels, and provided germplasm support for industrial development.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of edible fungus cultivation, and in particular relates to an excellent morel strain, a preparation method and application thereof. Background Art
[0002] In recent years, with the continuous development of the edible fungus industry, the scale of morel cultivation has also been expanding. However, due to the lack of high-yield morel strains, most planting bases have low yields or even no harvest each year, resulting in poor returns, which makes morel growers face greater planting risks.
[0003] Existing strains of morels generally have problems such as poor genetic stability and weak environmental adaptability, which makes it difficult to reproduce efficient cultivation. Related research on the life history of morels, such as artificial selection breeding, protoplast fusion, and hybrid breeding technology, has failed to make breakthrough progress.
[0004] Therefore, the selection and breeding of excellent Morchella strains and the preparation of fungus species are important tasks that need to be urgently addressed in Morchella research. Summary of the Invention
[0005] This study uses the Liumei Morel, a popular mushroom cultivated in Hanzhong, as experimental material. The spores are treated with atmospheric pressure room temperature plasma (ARTP) mutagenesis technology. The biological characteristics of the mutant strains at different growth stages (including mycelial growth characteristics, genetic characteristics, and variety comparison tests) are systematically analyzed to screen for high-yield Morel strains. This provides a theoretical basis and reference for the selection and breeding of superior Morel strains and the preparation of strains.
[0006] The invention provides a Morchellas extelata mutant strain M6SA24, with a preservation number of CCTCCNO: M 20251200.
[0007] The present invention also provides a cultivation method of a Morchella sextelata mutant strain M6SA24, characterized in that it comprises the following steps:
[0008] (1) Preparation of mother culture medium: The culture medium formula is as follows: 200.0 g potatoes, 20.0 g glucose, 5.00 g peptone, 5.00 g potassium dihydrogen phosphate, 3.00 g magnesium sulfate, 20.00 g agar, 10.00 mg vitamin B1, and 1000.00 mL distilled water per liter;
[0009] (2) Preparation of stock: The culture medium formula is: wheat 60%, rice husk 20%, wheat bran 18%, lime 1%, gypsum 1%;
[0010] (3) Preparation of cultivars: The culture medium formula is: wheat 60%, poplar wood chips 20%, humus 5%, fungus husk 8%, rice husk 5%, lime 1%, gypsum 1%;
[0011] (4) Nutritional pack: 68% wheat, 22% corn cobs, and 10% husks;
[0012] (5) Land preparation;
[0013] (6) Cover with film after row sowing to maintain the soil moisture content at 30% after sowing.
[0014] Furthermore, the specific steps of land preparation in step (5) are as follows: 15 days before sowing, 250 kilograms of quicklime are spread per mu, and deep rotary tillage and airing are performed until the soil particles are finely broken; the land is prepared to build a 1.2m wide × 0.2m high bed surface and dig a 0.3m wide × 0.2m deep drainage ditch; and three 10cm wide × 5cm deep sowing furrows are opened in the vertical direction.
[0015] Furthermore, the sowing amount of the drill sowing in step (6) is: 450 catties / mu.
[0016] The present invention further provides the fruiting body and mycelium of the mutant strain M6SA24 of Morchella sextelata.
[0017] The present invention further provides spores of the Morchella sextelata mutant strain M6SA24.
[0018] The present invention further provides a mushroom log of the Morchella sextelata mutant strain M6SA24.
[0019] The present invention further provides the use of the Morchella sextelata mutant strain M6SA24 in preparing fruiting bodies and / or mycelia.
[0020] Compared with the prior art, the present invention has the following beneficial effects:
[0021] This patent applies ARTP technology to the selection and breeding of superior strains of Morchella in the Qinba Mountain area for the first time. Through spore mutagenesis combined with phenotypic screening, superior Morchella strains with fast mycelial growth, early sclerotial formation and slow strain degeneration are obtained, providing germplasm support for breaking through the bottleneck of industrial development. This study used the six-sister Morchella, a major cultivated species in Hanzhong area, as experimental material, and used atmospheric pressure room temperature plasma (ARTP) mutagenesis technology to treat its spores. The biological characteristics of the mutant strains at different growth stages (mycelial growth characteristics, genetic characteristics and variety comparison tests, etc.) were systematically analyzed to screen and obtain high-yield Morchella strains. This provides a theoretical basis and reference for the selection and breeding of superior Morchella strains and strain preparation.
[0022] Description of biological deposit of Morchella sextelata mutant strain M6SA24:
[0023] Depository: China Center for Type Culture Collection;
[0024] Deposit number: CCTCC NO: M 20251200;
[0025] Deposit date: May 27, 2025;
[0026] Deposit address: Wuhan University, Wuhan, China;
[0027] Taxonomic name: Morchella esculenta. BRIEF DESCRIPTION OF THE DRAWINGS
[0028] Figure 1 This is a diagram showing the mycelial growth of the Morchella edible strain in Example 2;
[0029] Figure 2 This is a diagram showing the antagonistic reaction of the Morchella edodes strain in Example 2;
[0030] Figure 3 This is a microscopic morphology of mycelium of the Morchella mutant strain in Example 2;
[0031] Figure 4 Figure 2 is a microscopic structure diagram of the degenerated hyphae of the Morchella mutant strain in Example 2, wherein A1, B1, C1, and D1 represent the microscopic morphology of the first generation normal hyphae of the Morchella starting strain M6-2 and the mutant strains M6SA10, M6SA24, and M6SA25, respectively; A2, B2, C2, and D2 represent the microscopic morphology of the 13th generation degenerated hyphae of the Morchella starting strain M6-2 and the mutant strains M6SA10, M6SA24, and M6SA25, respectively;
[0032] Figure 5 These are the NBT staining results of the degenerated hyphae of the Morchella mutant strains in Example 2, where A1, B1, C1, and D1 represent the staining results of the first generation of hyphae of the Morchella starting strain M6-2 and the mutant strains M6SA10, M6SA24, and M6SA25, respectively; and A2, B2, C2, and D2 represent the staining results of the 13th generation of degenerated hyphae of the Morchella starting strain M6-2 and the mutant strains M6SA10, M6SA24, and M6SA25, respectively;
[0033] Figure 6 This is a statistical table of mycelial growth rates of different Morchella strains at different levels in Example 2;
[0034] Figure 7 Statistical diagram of agronomic traits of Morchella fruiting body in Example 2;
[0035] Figure 8 This is a diagram of the agronomic characteristics of some Morchella fruiting bodies in Example 2;
[0036] Figure 9 The fruiting conditions of each Morchella variety in Example 2 are shown. DETAILED DESCRIPTION
[0037] Example 1
[0038] 1. Preparation of spore suspension:
[0039] The fresh fruiting bodies of Morchella edulis with good mushroom shape, no pollution and strong body were put into a homogenizing bag, and 500ml sterile water was added to rinse the surface of the fruiting body. Then the suspension was mixed to obtain its ascospores, which were numbered as M6-2. The spore liquid was made into 10 by 10-fold concentration gradient dilution method. 6 CFU / mL suspension for mutagenesis
[0040] The Morchella siliquae numbered M6-2 was used as the test strain
[0041] 2. ARTP mutagenesis treatment:
[0042] The working gas is 99.9% high-purity helium, and the operating parameters are: power supply 120W, gas flow 10SLM, irradiation distance 2mm, and temperature <30℃.
[0043] The metal slide carried by the ARTP breeder was moved to the clean bench, and 10 μL of spore liquid was smeared on the metal slide, and then the plate was pushed into the mutagenesis chamber of the ARTP instrument. According to the operating procedures of the ARTP-IIS biological breeder, the power was set to 100 W, the air flow was set to 10 SLM, the distance between the slide and the air port was set to 3 mm, and the mutagenesis treatment time was set to: 0 s, 10 s, 20 s, 30 s, 40 s, 50 s, 60 s, 70 s, and 80 s.
[0044] After the mutagenesis was completed, the induced spores on the metal slide were washed with sterile saline to prepare a spore solution and vortexed evenly. The spores were counted using a hemocytometer to ensure that the number of spores was between 1×10 6 Each treatment was repeated three times, and the spores were cultured at 18°C. The spore growth was observed and the colony counts were performed on the plates. The strains were numbered M6SA1–M6SA n. The lethality of the strains was calculated according to the following formula to determine the optimal time for mutagenesis.
[0045] Lethality rate (%) = (total number of colonies in the control group - total number of colonies in the treatment group) / total number of colonies in the control group.
[0046] Generally, when the lethality rate is about 90%, there is a greater possibility of high-yield variation. Therefore, spores with a lethality rate of 90% are selected as test materials for screening mutant strains.
[0047] Transfer single colonies with a lethality rate above 90% to PDA medium and continuously isolate and purify them. Culture conditions: culture at 18°C until there are no contaminants in the plate, with 3 replicates for each sample.
[0048] Use the cross method to measure the hyphal growth rate, and record the hyphal density, hyphal color, sclerotium formation time, etc. Inoculate the purified strain onto a PDA slant medium, culture at 18°C for 7 days. When the hyphae cover the slant, transfer it to a refrigerator at 4°C for storage.
[0049] 3. Screening of the growth trend of mutant strains:
[0050] Transfer the Morchella mutant strains M6SA1–M6SA n obtained in step 2 to the mother culture medium, culture in a constant temperature incubator at 18°C for 7 days, with 3 replicates for each sample. Use the cross method to measure the hyphal growth rate, and record the hyphal density, hyphal color, sclerotium formation time, etc., and screen the strains with good growth conditions.
[0051] 4. Antagonism experiment:
[0052] Inoculate the mutant strains screened above and the original Morchella strain M6-2 in a "product" shape in the same petri dish, with an interval of about 2.5 cm between the strains. Culture in a constant temperature incubator at 18°C for 7 days, and observe the mutant strains with obvious antagonistic reactions between the mutant strains and the M6-2 strain.
[0053] 5. Observation of the microscopic structure of hyphae:
[0054] Transfer the strains screened in step 4 to the mother culture medium with a sterile punch with a diameter of 5 mm. Obliquely insert a sterile cover glass into the medium, insert 2-3 into each plate, culture at a constant temperature of 18°C. When the hyphae grow to cover about 1 / 2 of the cover glass, take out the cover glass and observe it under an optical microscope, and record the morphological characteristics of the hyphae.
[0055] 6. Determination of the hyphal lifespan of mutant strains:
[0056] The mycelial lifespan was determined by subculture. The prepared mother culture medium was divided into 500ml conical flasks and sterilized in an autoclave at 121℃ for 25min. Pour the culture medium into a 13cm×13cm square culture dish (about 30ml per dish) in a clean operating table, and after ultraviolet irradiation (wavelength 254nm) for 15-20min, use a 5mm diameter sterile puncher to inoculate the starting strain M6-2 and the mutant strain selected in step 5 at the top corner of the culture dish, and culture at a constant temperature of 18℃. When the mycelium grows to the other top corner of the diagonal of the culture dish, the sterile puncher repeats the above operation until the mycelium ages and dies, with 3 parallels for each strain. During this period, a portion of the culture medium was retained in the mother culture medium for each generation, and after being cultured at 18℃ until the slope was fully grown, it was transferred to a 4℃ refrigerator for storage and standby use. By recording the total culture time and total growth distance of each mutant strain and the starting strain throughout the process, the optimal mutant strain was selected for subsequent experiments.
[0057] 7. Observation of the microscopic morphology of degenerated strains of Morchella:
[0058] The microscopic morphology of mycelium was observed by using the insert method. The first and last generation mycelia of the strain screened in step 6 were taken for microscopic morphology observation, and the method was the same as that in step 6.
[0059] 8. NBT staining experiment:
[0060] In a clean bench, use a sterile punch with a diameter of 5 mm to inoculate each strain in the center of the mother culture medium. Insert the sterilized cover slip obliquely into the culture medium, insert 2-3 cover slips into each plate, and culture at 18°C. When the hyphae cover 1 / 2 of the cover slip, remove the cover slip and add 20 μl of 0.3 mmol·L -1 The NBT solution was dropped onto the dense hyphae area on the slide, reacted at room temperature for 25 min, and the hyphae color was observed under an optical microscope. This was repeated three times for each strain.
[0061] 9. Product comparison experiment:
[0062] The starting strain M6-2, mutant strain M6SA24, sixth sister morel N6, G8 and seventh sister morel No. 1 were selected for the experiment, and their mother species, original species and cultivated species were prepared respectively (the mother species was cultured at a constant temperature of 18°C; the original species and cultivated species were cultured at 18°C in the dark). The mother species culture medium contained 200.0g of potatoes, 20.0g of glucose, 5.00g of peptone, 5.00g of potassium dihydrogen phosphate, 3.00g of magnesium sulfate, 20.00g of agar, and 10.00g of vitamin B1 per liter. mg, 1000.00 mL of distilled water; the stock culture medium is: 60% wheat, 20% rice husks, 18% wheat bran, 1% lime, and 1% gypsum; the cultivated culture medium is: 60% wheat, 20% poplar wood chips, 5% humus, 8% fungus husks, 5% rice husks, 1% lime, and 1% gypsum; the nutrient bags (when the mycelium grows out of the soil surface, nutrient bags are placed, 4 per plot) are: 68% wheat, 22% corn cobs, and 10% rice husks.
[0063] The field experiment was conducted in Liuchuangou Village, Jiangkou Town, Liuba County, Hanzhong City. 15 days before sowing, 250 catties of quicklime was spread per mu and deep rotary tillage was carried out to dry the soil until the soil particles were fine. The land was prepared to build a 1.2 m wide × 0.2 m high bed and a 0.3 m wide × 0.2 m deep drainage ditch was dug. Three 10 cm wide × 5 cm deep sowing furrows were opened in the vertical direction. The seeds were sown in a row sowing method at an inoculum rate of 450 catties / mu. After sowing, the soil moisture content was maintained at about 30%, covered with black film, and a small arch shed was built to keep warm and moist, and ventilated regularly (each sample was about 1.44 m 2 , 5 replicates).
[0064] When the fruiting body changes from yellow-brown to dark brown and the cap ridges are distinct, the fruiting body is harvested. The fresh weight of each plot is weighed and calculated as g / 1.44m 2 Yield (mean ± SD) was calculated. Five fruiting bodies were randomly selected and the total length, cap diameter, length, and stipe diameter and length were measured using a vernier caliper. The data were statistically analyzed using SPSS 21.0.
[0065] Example 2
[0066] Selected samples: Morchella starting strain M6-2; Morchella mutant strain M6SA24; Main cultivated varieties of Morchella in Hanzhong area: Six-sister Morchella N6, G8; Seven-sister Morchella No. 1;
[0067] Analysis method: SPSS 21.0 statistical software was used to perform significance analysis of experimental data, and the numerical values in the results were expressed as mean ± standard deviation (AVE ± SD);
[0068] Use Excel 2019 and other related software to process images and charts. Verify the feasibility of the patent through growth rate measurement, antagonism experiments, microscopic observation, life span measurement, etc. of different Morchella edodes mutant strains;
[0069] Among them, the experimental process for verifying the feasibility of the patent is as follows:
[0070] 1. The mutant strains of Morchella obtained by mutagenesis were transferred to the mother culture medium and cultured in a constant temperature incubator at 18°C for 7 days. Each sample was replicated three times. The mycelial growth rate was measured using the cross-cross method, and the mycelial density, mycelial color, and sclerotia formation time were recorded to screen strains with good growth conditions.
[0071] The results are shown in Table 1 and Figure 1 As shown in the figure: Among the 45 mutant strains obtained by ARTP mutagenesis, 23 strains had better mycelial growth than the starting strain M6-2;
[0072] Among them, strains M6SA4, M6SA6, M6SA11, and M6SA12 showed the most significant increases in mycelial growth rate, increasing by 53.41%, 44.32%, 46.60%, and 45.45%, respectively, compared to M6-2. M6SA31, M6SA25, and M6SA23 showed slower increases, increasing by only 1.14%, 2.27%, and 3.41%, respectively. Sclerotia formation time also showed significant differences. Twelve strains, including M6SA6, M6SA11, M6SA12, M6SA17, and M6SA23, formed sclerotia in just 7 days, one day earlier than M6-2, while strains M6SA30 and M6SA31 required 10 days.
[0073] The morphological characteristics of some strains are as follows Figure 1 As shown, the number of sclerotia in strains M6SA4, M6SA31, M6SA23, and M6SA24 was significantly increased compared to strain M6-2. Among them, the sclerotia of M6SA4 were dispersed and clustered at the edges, while the sclerotia of the other three strains were evenly and densely distributed. In contrast, the number of sclerotia in strains M6SA6, M6SA11, M6SA12, and M6SA25 was relatively low. In summary, ARTP mutagenesis significantly altered the mycelial growth characteristics of the strains.
[0074] Table 1 Mycelial growth record of Morchella mutant strains
[0075]
[0076]
[0077] Note: + indicates the degree of growth, and the more +, the better the growth. The data in the table are expressed as mean ± standard error; "*" indicates significant difference (p < 0.05), and "**" indicates extremely significant difference (p < 0.01)
[0078] 2. The mutant strains screened in step 1 were inoculated in the same dish with the starting strain M6-2 in a "pin" pattern, with an interval of about 2.5 cm between the strains. The plates were placed in a constant temperature incubator at 18°C for 7 days to observe the mutant strains that showed obvious antagonistic reactions with the M6-2 strain.
[0079] The results are as follows Figure 2 As shown: A total of 10 mutant strains were obtained that had obvious antagonistic reactions with the starting strain M6-2, and the strains that had obvious antagonistic reactions with the starting strain were M6SA10, M6SA11, M6SA12, M6SA23, M6SA24, M6SA25, M6SA27, M6SA28, M6SA29 and M6SA31.
[0080] 3. Transfer the strain selected in step 2 to the mother culture medium using a sterile hole punch with a diameter of 5 mm. Insert sterile coverslips obliquely into the culture medium, inserting 2-3 per plate. Incubate at a constant temperature of 18°C. When the mycelium grows to cover about 1 / 2 of the coverslip, remove the coverslip, observe under an optical microscope, and record the mycelial morphology.
[0081] like Figure 3 As shown: A total of 6 mutant strains were screened, namely M6SA10, M6SA11, M6SA12, M6SA24, M6SA25 and M6SA28. Secondly, the angles between the hyphae and the main hyphae are mostly acute, and the hyphae edges are neat and have many branches.
[0082] 4. The mycelial life span was determined by subculture. The prepared mother culture medium was divided into 500 ml conical flasks and sterilized in an autoclave at 121°C for 25 minutes.
[0083] Pour the solution into a 13 cm x 13 cm square Petri dish (approximately 30 ml per dish) in a cleanroom. After UV irradiation for 15-20 minutes, use a 5 mm diameter sterile hole punch to inoculate the starting strain M6-2 and the mutant strain selected in 3.3.1.3 at the top corner of the dish. Incubate at 18°C.
[0084] When the mycelium grows to the other diagonal corner of the culture dish, repeat the above operation with a sterile punch until the mycelium ages and dies. Each strain should be replicated three times.
[0085] During each generation, retain a portion of the culture in the mother culture medium and cultivate at 18°C until the slant is fully grown. Afterwards, transfer the culture to a 4°C refrigerator for future use. By recording the total cultivation time and total growth distance of each mutant strain compared to the starting strain, the optimal mutant strain was selected for subsequent experiments.
[0086] As shown in Table 2 : The mycelial lifespans of mutant strains M6SA11, M6SA12, and M6SA28 were all shorter than those of the starting strain M6-2 (growth 3125.33 h, mycelial length 186.20 cm), and all three died at the 12th generation;
[0087] In contrast, M6SA10, M6SA24, and M6SA25 showed better survival ability: M6SA10 (3453.67 h) and M6SA24 (3489.00 h) survived to the 14th generation, and M6SA25 (3202.00 h) survived to the 13th generation. Their total growth time was significantly longer than that of the starting strain.
[0088] The starting strain M6-2 still maintained a small amount of sclerotia formation at the 12th generation, but showed significant edge irregularity and pigment accumulation at the 13th generation.
[0089] Among the mutant strains, M6SA10 and M6SA25 completely lost their ability to form sclerotia by the 12th generation, but developed well-developed aerial hyphae. In contrast, M6SA24 retained a small number of sclerotia at the 13th generation and exhibited the lowest pigment accumulation. Based on a comprehensive evaluation of hyphal lifespan, sclerotial formation, and pigment accumulation, three mutant strains, M6SA10, M6SA24, and M6SA25, were selected for further studies, demonstrating long lifespan (>3200 hours), high sclerotial viability, and low pigmentation.
[0090] Table 2 Mycelial lifespan record of Morchella mutant strains
[0091]
[0092] 5. Use the insert method to observe the mycelial micromorphology. Take the first and last generation mycelia of the strain screened in step 4 for microscopic morphology observation. The method is the same as step 3.
[0093] The results are as follows Figure 4 As shown in the figure: the first generation hyphae structure of the starting strain M6-2 and the mutant strains M6SA10, M6SA24, and M6SA25 were more obvious, with longer hyphae and clear boundaries. The angles between the main and secondary hyphae were mostly acute. However, the secondary hyphae of the 13th generation hyphae after subculture were shorter, the hyphae crossed each other more, and the angle between the main and secondary hyphae increased.
[0094] 6. In a clean bench, use a sterile punch with a diameter of 5 mm to inoculate each strain in the center of the mother culture medium. Insert the sterilized cover slip obliquely into the culture medium. Insert 2-3 cover slips into each plate. Incubate at 18°C. When the hyphae cover 1 / 2 of the cover slip, remove the cover slip and add 20 μl of 0.3 mmol·L -1Drop the NBT solution onto the dense hyphae area on the slide, let it react at room temperature for 25 minutes, and then observe the hyphae color under an optical microscope. Repeat this process three times for each strain.
[0095] The results are as follows Figure 5 As shown, the first-generation strains (A1, B1, C1, and D1) initially stained dark blue-purple, which faded to light purple after the reaction. In contrast, the 13th-generation strains (A2, B2, C2, and D2) retained a dark blue-purple color after staining. Compared to the 13th-generation samples, C2 (M6SA24) stained the lightest. These results indicate that the ROS-scavenging ability of all 13th-generation strains was weaker than that of the first generation, confirming that subculture induces mycelial degeneration, with mutant strain M6SA24 (C2) exhibiting the lowest degree of degeneration.
[0096] 7. The starting strain M6-2 of Morchella, the mutant strain M6SA24, the sixth sister Morchella N6, G8 and the seventh sister Morchella No. 1 were selected for the experiment, and their mother species, original species and cultivated species were prepared respectively. The mother species culture medium was 200.0g of potato, 20.0g of glucose, 5.00g of peptone, 5.00g of potassium dihydrogen phosphate, 3.00g of magnesium sulfate, 20.00g of agar, 10.00mg of vitamin B1, and 1000.00mL of distilled water; the original species culture medium was 60% wheat, 20% rice husk, 18% wheat bran, 1% lime, and 1% gypsum; the cultivated species culture medium was 60% wheat, 20% poplar wood chips, 5% humus soil, 8% fungus bran, 5% rice husk, 1% lime, and 1% gypsum; the nutrient pack was 68% wheat, 22% corn cobs, and 10% rice husk. The field test was conducted in Liuchuangou Village, Jiangkou Town, Liuba County, Hanzhong City. 250 catties of quicklime was spread per mu 15 days before sowing, and deep rotary tillage and drying were carried out until the soil particles were fine. The land was prepared to build a 1.2m wide × 0.2m high bed surface and a 0.3m wide × 0.2m deep drainage ditch was dug. Three 10cm wide × 5cm deep sowing furrows were opened in the vertical direction. The seeds were sown in strips at an inoculation rate of 450 catties / mu. After sowing, the soil moisture content was maintained at about 30%. In the early growth stage: growth began at a low temperature of 1-2°C. If the temperature exceeded 28°C, the growth would slow down or stop. Fruiting body development period: the temperature is required to be 4-16°C, and the most suitable is 8-12°C. Cover with black film, build a small arch shed, keep warm and moist, and ventilate regularly (each sample plot is about 1.44m 2 , 5 replicates). The fruiting bodies were harvested when their color changed from yellow-brown to dark brown and the cap ridges were distinct. The fresh weight of each plot was weighed and calculated as g / 1.44m 2 Yield (mean ± SD), 5 fruiting bodies were randomly selected and the total length, cap diameter, length, and stipe diameter and length were measured with a vernier caliper, and the data were statistically analyzed using SPSS 21.0;
[0097] The results are as follows Figure 6-9As shown in the figure: In the mother culture medium, the mycelial growth rate of M6SA24 reached 1.18±0.08acm / d, which was 35.63% higher than that of the starting strain M6-2 (0.87±0.06c cm / d), and dense sclerotia were formed as early as 7 days (M6-2 required 8 days). Morchella N6 grew the slowest (0.83±0.06c cm / d).
[0098] In the stock culture stage, M6SA24 maintained its dominance, with a hyphal growth rate of 1.40±0.10a cm / d (25% higher than M6-2) and sclerotia formation in 9 days, while M6-2 and other strains (except N6) required 10 days, and Morchella N6 required the latest 11 days. In the cultivar culture medium, M6SA24 led with a growth rate of 1.24±0.09a cm / d (6.90% higher than M6-2), forming sclerotia in 10 days, while Morchella N6 (1.04±0.05b cm / d) and G8 both required 11 days.
[0099] The mycelia appeared pure white at all stages, and the mutant strain M6SA24 was significantly superior to the control in terms of mycelial growth rate (mother strain +35.63%, original strain +25%, cultivated strain +6.90%) and sclerotia formation time (2 days earlier, 1 day earlier, and at the same time, respectively).
[0100] Each morel strain is at 1.44m 2 The fruiting performance of the samples showed significant differences. In terms of fruiting density, the M6SA24 strain had a fruiting density of 58.8±1.10 a / 1.44 m 2 The value of the strain M6-2 ranked first, which was significantly higher than 51.8±0.75b / 1.44m 2 , while the lowest density of strain G8 was only 22.2±1.10e / 1.44m 2 Compared with M6-2, Morchella N6, No. 1 and G8 strains, the fruiting density of M6SA24 increased by 13.51%, 66.10%, 89.69% and 164.86% respectively;
[0101] The fruiting body morphology of the Morchella strains showed significant differentiation, with mutant strain M6SA24 exhibiting the best agronomic traits. Its total fruiting body length reached 15.10±0.26 cm, a 25.83% increase compared to the original strain M6-2. Specifically, the cap length reached 10.14±0.40 cm (accounting for 67.15% of the total length), the stipe length was 5.00±0.19 cm, and the cap diameter was 3.70±0.23 cm.
[0102] Comparative analysis showed that the fruiting body length of M6SA24 was 7.09% longer than Morchella No. 1 (14.10 ± 0.55 cm), 13.53% longer than Morchella N6, and 31.30% longer than G8. Notably, this strain exhibited particularly strong cap development, with its cap being 2.03 times longer than the stipe, demonstrating a significant apical growth advantage.
[0103] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.
Claims
1. A mutant strain M6SA24 of Morchella sextelata, with a deposit number of CCTCC NO: M20251200.
2. A cultivation method for the Morchella sextelata mutant strain M6SA24 according to claim 1, characterized in that: The following steps are involved: (1) Preparation of mother culture: The culture medium formula is as follows: 200.0 g potatoes, 20.0 g glucose, 5.00 g peptone, 5.00 g potassium dihydrogen phosphate, 3.00 g magnesium sulfate, 20.00 g agar, 10.00 mg vitamin B1, and 1000.00 mL distilled water; (2) Preparation of stock: The culture medium formula is: wheat 60%, rice husk 20%, wheat bran 18%, lime 1%, gypsum 1%; (3) Preparation of cultivars: The culture medium formula is: wheat 60%, poplar wood chips 20%, humus 5%, fungus husk 8%, rice husk 5%, lime 1%, gypsum 1%; (4) The nutrient pack consists of 68% wheat, 22% corn cobs, and 10% husks; (5) Land preparation; (6) Cover with film after row sowing to maintain the soil moisture content at 30% after sowing.
3. The cultivation method according to claim 2, wherein The specific steps of land preparation in step (5) are as follows: 15 days before sowing, 250 kilograms of quicklime are spread per mu, and deep rotary tillage and airing are performed until the soil particles are finely broken; the land is prepared to build a 1.2m wide × 0.2m high bed surface and dig a 0.3m wide × 0.2m deep drainage ditch; and three 10cm wide × 5cm deep sowing furrows are opened in the vertical direction.
4. The cultivation method according to claim 2, wherein The sowing amount of the drill sowing in step (6) is: 450 catties / mu.
5. The fruiting body and mycelium of the Morchella sextelata mutant strain M6SA24 according to claim 1.
6. Spores of the Morchella sextelata mutant strain M6SA24 according to claim 1.
7. A mushroom log containing the Morchella sextelata mutant strain M6SA24 according to claim 1.
8. Use of the Morchella sextelata mutant strain M6SA24 according to claim 1 in preparing fruiting bodies and / or mycelia.