High-COtolerance pleurotus pulmonarius variety suitable for one-time harvesting and application of pleurotus pulmonarius variety

Abstract

The invention discloses a high-CO2-tolerance pleurotus pulmonarius variety suitable for one-time harvesting and application of the pleurotus pulmonarius variety. The pleurotus pulmonarius variety is excellent in character, meets the market diversified requirements, is suitable for industrial cultivation and has good application and popularization prospects. The Pleurotus pulmonarius variety 'Shenxiu No.2' has high CO2 tolerance, and can tolerate 10000 ppm of high-concentration CO2 in the bag opening and fruiting period; the early-maturing characteristic is achieved, about 10 days shorter than that of an existing main cultivated variety, and the cultivation period is short; the method has the advantages of being convenient to operate, labor-saving and time-saving, fruiting is concentrated, uniformity is good, and one-time harvesting and packaging can be achieved; the ISSR molecular marker combination has the characteristic of high yield, the average first-tide yield of each bag is 198.25 / g, the sporocarp pileus of the pleurotus pulmonarius variety 'Shenxiu No.2' is light brown and is slightly large, and the ISSR molecular marker combination of the pleurotus pulmonarius variety 'Shenxiu No.2' has specificity and specificity for identifying the pleurotus pulmonarius variety 'Shenxiu No.2'.

Description

Technical Field

[0001] The invention belongs to the technical field of Pleurotus pulmonaria breeding and strain molecular identification, and particularly relates to a Pleurotus pulmonaria variety that is tolerant to high CO2 and suitable for one-time harvesting and an application thereof. Background Art

[0002] Pleurotus pulmonarius belongs to the Fungi, Basidiomycetes, Agaricales, Pleurotaceae, and Pleurotus genus. It typically grows on trees such as oaks, elms, and maples in temperate regions, as well as on fallen branches, dead and decaying stumps, and wet wood. Its fruiting bodies are beautifully shaped, tender, crisp, and delicious, making them highly sought after by consumers. They are also rich in nutrients such as protein, crude fat, polysaccharides, and various vitamins. The mycelium and fruiting bodies are also rich in active substances such as polysaccharides, which have antioxidant, anti-tumor, lipid-lowering, anti-mutagenic, and immune-boosting properties.

[0003] Currently, the main production areas of Pleurotus pulmonaria primarily utilize seasonal bagged cultivation in greenhouses. While this method offers low investment and high yields (seven harvest waves), it also faces challenges such as year-round availability of fresh mushrooms, unstable yields, and frequent pest and disease outbreaks. In recent years, industrialized cultivation of Pleurotus pulmonaria has rapidly developed, offering advantages such as year-round supply of fresh mushrooms, stable yield and quality, and manageable pests and diseases. This has been particularly evident due to the unprofitability of bagged King Oyster Mushroom cultivation, the shift to traditional cultivars such as edible mushroom shelters, and the introduction of new cultivation methods. However, the lack of specialized varieties for industrialized production of Pleurotus pulmonaria and varieties with high CO2 tolerance has hindered the current trend of industrialized Pleurotus pulmonaria production. Furthermore, the primary varieties used for Pleurotus pulmonaria production are 'Jinxiu' and 'Taixiu' varieties, with a production cycle (from inoculation to harvest) of approximately 50 days (43-44 days for incubation and 6-7 days for fruiting). Harvesting, particularly with the "large, small" root pruning and grading method, results in high labor costs. Summary of the Invention

[0004] The purpose of the present invention is to provide a high CO2 tolerant Pleurotus pulmonaria variety 'Shenxiu No. 2' and its application, which has high CO2 tolerance, concentrated buds and a large number of effective stems, in response to the phenomenon that the number of effective buds is small in the existing production of Pleurotus pulmonaria.

[0005] The purpose of the present invention is to solve the problems of long harvesting time and high labor consumption in the existing production of Pleurotus pulmonaria, and to provide a Pleurotus pulmonaria variety 'Shenxiu No. 2' with good bud uniformity, dense and hard texture, suitable for one-time harvesting and packaging, and its application.

[0006] The purpose of the present invention is to provide a Pleurotus pulmonaria variety 'Shenxiu No. 2' with a cultivation period 10 days shorter than that of the main cultivated varieties and its application in view of the current breeding status of Pleurotus pulmonaria and the long production period of the main cultivated varieties.

[0007] The purpose of the present invention is to provide a Pleurotus pulmonaria variety 'Shenxiu No. 2' with a funnel-shaped fruiting body cap cross-section and light brown color, and its application, in response to the current consumption status of Pleurotus pulmonaria and the phenomenon that the fruiting body caps of the main cultivated varieties are mostly black or brown and relatively single in color.

[0008] The object of the present invention is to provide a factory cultivation method for the above-mentioned Pleurotus pulmonaria variety.

[0009] Another object of the present invention is to provide a molecular identification method for the above-mentioned Pleurotus pulmonaria variety.

[0010] The Pleurotus pulmonarius 'Shenxiu No. 2' variety of the present invention was deposited on March 7, 2025 in the Guangdong Provincial Microbial Culture Collection Center, with the address being: 5th Floor, Building 59, Compound 100, Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, with the deposit number being: GDMCC No: 65787.

[0011] The aforementioned Pleurotus pulmonaria cultivar 'Shenxiu No. 2' was derived from a wild strain of Pleurotus pulmonaria collected in Shigatse City, Tibet Autonomous Region. The strain was isolated and purified through tissue isolation. The strain was then systematically domesticated and selected through small-scale, pilot, and demonstration cultivation. Mycelium of 'Shenxiu No. 2' grown on standard PDA medium exhibits moderate growth, is dense, and lacks aerial hyphae. The colonies are round and lack pigment. The optimal growth temperature for mycelium is 25°C to 28°C, with an average growth rate of 4.23±0.03 mm / day at 25°C. The fruiting bodies are clustered, with lateral stipes. The buds are initially light brown, with numerous effective stems and a dense, firm texture. The optimal fruiting temperature is 20°C to 24°C, and the CO2 concentration during the fruiting period should be controlled between 3000ppm and 10000ppm. The cross-section of the mature fruiting body's cap is funnel-shaped and light brown in color. The fruiting body produces mushrooms in a relatively uniform pattern, with a large average number of effective stems. The overall commercial properties and quality are good, and the fruiting body can be harvested and packaged in one go, making it a variety suitable for factory cultivation.

[0012] The molecular identification method for the aforementioned Pleurotus pulmonate variety ‘Shenxiu 2’ involves six ISSR primers, as follows:

[0013] ISSR 1: 5′-TGCACACACACACAC-3′;

[0014] ISSR 2: 5′-GTGACACACACAC-3′;

[0015] ISSR 3: 5′-GTGACGACTCTTCTCCTCTCT-3′;

[0016] ISSR 4: 5′-GGATGCAACACACACACAC-3′;

[0017] ISSR 5: 5′-CGTGTGTGTGTGTGT-3′;

[0018] ISSR 6: 5′-AGTGTGTGTGTGTGT-3′.

[0019] Beneficial effects of the present invention: The present invention uses a wild Pleurotus pulmonaria strain collected in the Tibet Autonomous Region as the starting material, and through tissue separation, small-scale trials, pilot tests and demonstration application cultivation, systematically domesticates and selects a new variety 'Shenxiu No. 2' with high CO2 tolerance, suitable for one-time harvesting, early maturity, high yield and suitability for factory cultivation.

[0020] The factory-cultivated Pleurotus pulmonaria cultivar 'Shenxiu No. 2' maintains a CO2 concentration of around 3500 ppm or below during the mycelial culture period; 5500-6000 ppm during the ripening period; and 3000-10000 ppm during the bagging and fruiting period. The average yield of the first-batch mushrooms per bag (17 cm × 35 cm × 0.045 cm) reaches 197.63 g / bag, a 10.88% increase over the 178.24 g / bag of the primary control cultivar 'Taixiu'. The cultivation cycle is 10 days shorter than that of the primary control cultivar 'Taixiu'. 'Shenxiu No. 2' has a light brown cap with a funnel-shaped cross-section, averaging 6.47 cm in length, 6.07 cm in width, and 0.63 cm in thickness. The stipe is white, averaging 4.74 cm in length and 1.17 cm in diameter. The fruiting body has a large number of effective stems and a dense, firm texture. 'Shenxiu 2' has excellent traits, meeting the requirements of factory cultivation for early maturity, high yield, and labor-savings, while also meeting the diverse market demands. It is suitable for factory cultivation and has good prospects for application and promotion. The ISSR molecular marker method for the Pleurotus pulmonaria 'Shenxiu 2' offers advantages such as high reproducibility, high polymorphism, and low cost, and is highly specific and specific for identifying the Pleurotus pulmonaria strain 'Shenxiu 2'. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments, wherein:

[0022] Figure 1 This is the phylogenetic tree diagram of ‘Shenxiu 2’.

[0023] Figure 2This is the antagonism diagram between 'Shenxiu 2' and the main production variety 'Taixiu', as well as the commonly used strains 'Shenxiu 1' and 'Dongxiu'.

[0024] Figure 3 The electrophoresis diagram of different Pleurotus pulmonaria strains amplified using ISSR primers 1 to 6, where M is a DNA molecular weight standard, 1 is '3108', 2 is 'Shenxiu No. 1', 3 is 'Taixiu', 4 is 'Jinxiu', 5 is 'Shenxiu No. 2', 6 is 'Dongxiu', and 7 is 'SX3'.

[0025] Figure 4 This is the phylogenetic analysis diagram of 7 Pleurotus pulmonaria strains based on the unweighted arithmetic mean pairing method, where 1 is '3108', 2 is 'Shenxiu 1', 3 is 'Taixiu', 4 is 'Jinxiu', 5 is 'Shenxiu 2', 6 is 'Dongxiu', and 7 is 'SX3'.

[0026] Figure 5 The fruiting body morphology of 'Shenxiu No. 2' (left) and the main production variety 'Taixiu' (right).

[0027] Figure 6 Illustration of the harvesting and packaging of Shenxiu No. 2 (top) and the main production variety Taixiu (bottom). DETAILED DESCRIPTION

[0028] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with specific embodiments.

[0029] Example 1:

[0030] Breeding of the Pleurotus pulmonaria cultivar 'Shenxiu No. 2': Wild Pleurotus pulmonaria fruiting bodies collected in the Tibet Autonomous Region were isolated and purified using tissue separation. The specific method is as follows: Dirt, weeds, and other impurities were removed from the fruiting bodies. After placing them in a clean bench, the surface was wiped with 75% alcohol cotton and then wiped again with sterile water. The surface moisture was then absorbed and placed in a Petri dish. Using a sterile scalpel, the stem or stipe tissue inside the fruiting body was cut into 1×1.5 cm pieces. These pieces were then inoculated with an inoculating needle into the center of a 90 mm diameter Petri dish containing 20 mL of potato dextrose agar (PDA) medium. The fruiting bodies were then cultured in a dark incubator at 25°C to obtain the Pleurotus pulmonaria strain. The tip hyphae of the 5-day-old culture were picked and transferred to a new PDA medium for purification. After 5 purifications, the best-growing strain '0379' was selected based on the mycelial growth rate, colony morphology, mycelial morphology under microscopic examination, and the number of lock-like joints. It was inoculated into a test tube containing PDA medium for culture. After the mycelium was fully grown, it was placed in a 4°C storage room for storage until use.

[0031] The mycelial growth rate test of the strain '0379' that was successfully isolated from the tissue was conducted. The activated bacterial block was taken with a 5mm diameter puncher and inoculated into the center of a culture dish containing PDA culture medium, and cultured at a constant temperature of 25°C in the dark. The experiment was repeated three times, and the mycelial growth rate and growth potential were recorded. The average growth rate of the strain '0379' on PDA culture medium was 4.23mm / d, which is comparable to the average growth rate of the main cultivated variety 'Taixiu' on PDA culture medium (4.26mm / d). The mycelial growth of the strain '0379' was medium, the mycelium was relatively dense, there were no aerial mycelium, and the colony shape was relatively round.

[0032] The strain '0379' with successful components was domesticated and cultivated in a small trial. 50 production bags were set up at each site for domestication and 300 production bags were set up at each site for small trial. The conventional substitute material formula (30% corn cobs, 25% sawdust, 13% bran, 13% rice bran, 7% cottonseed hulls, 7% soybean meal, 3% corn flour, 1% lime, 1% light calcium carbonate, with a water content of about 67%) was used for cultivation. The materials were packed in polyethylene plastic bags of 17cm×35cm×0.045cm. Each bag contained 1.1kg~1.2kg of wet material and about 0.45kg of dry material. The material height was 18cm~20cm and 1.5×10 5 After inoculation, the bags were placed in a culture room at 23-25°C for incubation in the dark.

[0033] After the mycelium has developed to full bags, the mushrooms are transferred to the fruiting room for fruiting management, and fruiting characteristics are recorded and counted. The first-wave mushroom yield of strain '0379' averaged 167.58g per bag, with a biological conversion efficiency of 37.24%, 0.32% higher than the first-wave yield of the main cultivated variety 'Taixiu' (167.05g per bag). The growth cycle of strain '0379' is approximately 40 days, 10 days shorter than that of the current main cultivated variety 'Taixiu'. The young buds of strain '0379' are light brown, and the fruiting bodies have light brown caps, white, slender stipes, a firm texture, and uniform fruiting. Fruiting bodies with excellent fruiting were selected for further tissue separation and coded into different series for the aforementioned fruiting tests. The series with excellent fruiting performance was selected for subsequent pilot tests.

[0034] Strain '0379' was cultivated in a pilot-scale test. Each pilot-scale test site was equipped with 800 production bags. The cultivation formula was the same as the pilot test, and cultivation techniques were tailored to the strain's characteristics and commercial requirements. The main cultivated Pleurotus pulmonaria cultivar 'Taixiu' was used as a control. Agronomic data, including growth cycle, average first-bush yield per bag, and number of effective buds, are shown in Table 1. The results showed that '0379' exhibited a short growth cycle for the first-bush mushrooms, high yield, and concentrated and uniform fruiting, making it a suitable variety for industrial cultivation.

[0035] Table 1 Agronomic characteristics of the pilot strain '0379'

[0036]

[0037]

[0038] Strain '0379' was further cultivated in pilot and demonstration trials to refine cultivation techniques, using the dominant cultivar 'Taixiu' as a control. The results (Table 2) showed that strain '0379' had a higher average first-wave yield than the dominant cultivar, a 10-day shorter cultivation cycle, and a concentrated and uniform budding pattern, making it suitable for single-stage harvesting and packaging. This strain met the requirements for early maturity, high yield, and labor-saving factory cultivation. The strain was ultimately named 'Shenxiu No. 2'.

[0039] Table 2 Comparison of agronomic traits between the new strain ‘Shenxiu 2’ and the control variety ‘Taixiu’

[0040] strain Shin Soo No. 2 Taiwan Show Average yield of first-wave mushrooms per bag / g 197.63 178.24 Culture cycle / d 40 50 Number of effective buds / piece 82 40 Cap width / cm 6.07 4.85 Cap thickness / cm 0.63 0.50 Cap length / cm 6.47 5.70 Cross-sectional morphology of the cap funnel Shallow funnel (concave) Cap color light brown brown Stem diameter / cm 1.17 1.81 Stem length / cm 4.74 5.52 Stem color White White Stem texture Harder soft Bud color light brown brown

[0041] Example 2:

[0042] Molecular biological identification of the Pleurotus pulmonate variety 'Shenxiu 2':

[0043] A high-specificity PCR amplification kit (Shanghai Sangon Biotech Co., Ltd.) was used to extract DNA from strain ‘Shenxiu No. 2’. The DNA purity and concentration were detected by 1.2% agarose gel electrophoresis and a micro-UV-visible spectrophotometer, respectively. The DNA concentration of strain ‘Shenxiu No. 2’ was adjusted to 50 ng / μL and stored in a −20°C refrigerator until use.

[0044] PCR amplification was performed using ITS1 and ITS4 as primers, and the amplified products were sequenced. The primer sequences are as follows:

[0045] ITS1: 5′-TCCGTAGGTGAACCTGCGG-3′;

[0046] ITS 4: 5′-TCCTCCGCTTATTGATATGC-3′.

[0047] The ITS-PCR reaction system is as follows: total volume 20 μL: 1 μL DNA template (concentration 50 ng / μL), 24 μL 10x Buffer (containing Mg 2+ ), 0.5 μL of 2.5 mM dNTP, 0.2 μL of 5 U / μL DNA polymerase, 0.5 μL of 0.2 μM ITS1 primer, 0.5 μL of 0.2 μM ITS4 primer, and ddH2O to 20 μL.

[0048] The ITS-PCR reaction conditions were as follows: initial denaturation at 94°C for 5 minutes, 30 cycles of 94°C for 1 minute, 60°C for 1 minute, and 72°C for 75 seconds, followed by extension at 72°C for 10 minutes. PCR products were detected using 1.2% agarose gel electrophoresis.

[0049] The PCR product was sent to Kangwei Century Biotechnology Co., Ltd. for sequencing. The nucleotide sequence of the PCR product was shown in SEQ ID No: 1 (ITS sequence).

[0050] The ITS sequence of the strain 'Shenxiu No. 2' of the present invention was BLAST-compared to the NCBI nucleic acid database. Sequences with high homology were selected for comparison analysis, and a phylogenetic tree was constructed using the maximum likelihood method of MEGA11. The results showed that the strain 'Shenxiu No. 2' of the present invention had the highest similarity with Pleurotus pulmonarius, with a similarity of up to 99.85%. The phylogenetic tree is shown in Figure 1. Figure 1 The results show that the strain 'Shenxiu No. 2' of the present invention belongs to the genus Pleurotus pulmonarius in terms of classification.

[0051] Shenxiu 2 ITS sequencing sequence:

[0052]

[0053]

[0054] Example 3:

[0055] Genetic characteristics of the Pleurotus pulmonate variety ‘Shenxiu No. 2’:

[0056] (1) Antagonistic reaction

[0057] According to the agricultural industry standard "Antagonistic reaction of differentiating edible fungi" (NY / T1845-2010), antagonistic experiments were conducted and it was found that obvious ridge-type and groove-type antagonistic reactions could be observed between 'Shenxiu No. 2' and commonly used varieties on the market ('Taixiu' and 'Shenxiu No. 1') and a wild strain ('SX3'). Figure 2 ), indicating that the new strain 'Shenxiu No. 2' is a different species from other strains.

[0058] (2) Comparative analysis of ISSR molecular markers

[0059] The agricultural industry standard "ISSR Method for Authenticity Identification of Edible Fungi" (NY / T1730-2009) was used to identify the differential identity of 'Shenxiu No. 2', commercially available cultivars 'Taixiu', 'Jinxiu', 'Dongxiu', and '3108', as well as the independently bred 'Shenxiu No. 1' and the wild strain 'SX3'. PCR amplification was performed using 20 ISSR primers (Table 3).

[0060] Table 3 ISSR primers and their sequences

[0061]

[0062]

[0063] PCR amplification system (10 μL): 1 μL 10× PCR buffer, 0.8 μL dNTP (2.5 mmol / L), 0.1 μL HSTaq DNA enzyme (5 U / μL), 1.2 μL ISSR primer (5 μmol / L), 1 μL template DNA (20-30 ng / μL), 5.9 μL ddH2O.

[0064] The PCR reaction procedure was as follows: 95°C pre-denaturation for 5 minutes; 35 cycles of 95°C denaturation for 30 seconds, 60°C annealing for 30 seconds, and 72°C extension for 30 seconds; and 60°C extension for 30 minutes. PCR products were detected using 1.2% agarose gel electrophoresis, and product sizes were recorded. Electrophoresis results showed that primers ISSR1 to ISSR6 could effectively identify significant differences between 'Shenxiu No. 2' and other strains ( Figure 3 and Figure 4 ), ‘Shenxiu No. 2’ is a different variety of Pleurotus ostreatus with a lung shape.

[0065] Example 4:

[0066] Factory cultivation method of the lung-shaped Pleurotus eryngii variety 'Shenxiu No. 2':

[0067] (1) Stock culture: The strain 'Shenxiu No. 2' was inoculated on the stock culture medium and cultured in the dark for 8 days at a temperature of 25°C and a relative humidity of 70%. The stock culture medium is: 200g potato, 20g glucose, 20g agar, natural pH, sterile water to 1000mL, 121°C, 1×10 5 Sterilize Pa for 20 minutes, pour into a 90 mm diameter Petri dish, and cool for later use.

[0068] (2) Stock culture: The culture medium cultured on the culture dish in step (1) is cut into 1cm×1cm mycelium blocks, inoculated on the stock culture medium for culture, and half a culture dish of mycelium blocks are inoculated in each bag. The culture is cultured in the dark at a temperature of 23℃~25℃ and a relative humidity of 65%~70% for 30~35 days. The stock culture is obtained after the mycelium fills the bag. The stock culture medium formula is: 30% corn cobs, 25% sawdust, 13% bran, 13% rice bran, 7% cottonseed hulls, 7% soybean meal, 3% corn flour, 1% lime, 1% light calcium carbonate, with a water content of about 67% and a pH value of 7.0-8.0. The stock culture medium preparation method is: various raw materials are mixed and stirred for 30 minutes, and then water is added according to a material-water ratio of 1:1.1~1.3 and stirred for 30 minutes. A polyethylene plastic bag with a specification of 17cm×35cm×0.045cm is selected. The culture medium is placed in a cultivation bag, with each bag containing 1.1kg to 1.2kg of wet material and about 0.45kg of dry material, with a height of 18cm to 20cm. After filling, it is sealed with a plastic ring, plugged with cotton, and sterilized by autoclaving at 126℃ for 4 hours.

[0069] (3) Cultivation of cultivars: The original seed of 'Shenxiu No. 2' obtained in step (2) is inoculated onto a cultivar culture medium at a mass ratio of 1:20 to 1:30. Cultivation is carried out at a temperature of 23°C to 25°C and a relative humidity of 65% to 70% in the dark. Cultivation is performed until the mycelium fills the bag, thereby obtaining the cultivar. The cultivar culture medium is formulated and prepared in the same manner as the original seed culture medium in step (2).

[0070] (4) Inoculation: In a sterile inoculation room or sterile environment, the 'Shenxiu No. 2' cultivar obtained in step (3) is inoculated into the rod hole in the center of the cultivation bag at a mass ratio of 1:20 to 1:30, and then sealed with a collar and a sealing plug. The cultivation is carried out at a temperature of 23°C to 25°C, a relative humidity of 65% to 70%, and a CO2 concentration of approximately 3500 ppm or less, in the dark, until the mycelium fills the bag. The culture material contained in the cultivation bag is the same as the original seed culture medium formula and preparation method described in step (2), but a 13 cm hollow plastic rod is inserted into the center of the cultivation bag during bagging.

[0071] (5) After-ripening: After the mycelium has grown full in the bag, the temperature is controlled at 26℃~28℃ and the CO2 concentration is controlled at 5500ppm~6000ppm. After-ripening culture is carried out for about 10 days to achieve physiological maturity.

[0072] (6) Scraping the mycelium: After the mycelium culture is completed, move it into the fruiting room. Cut the film off the bag along the surface of the material from step (5), open the bag to expose the material surface, and use a sterilized knife to scrape off the surface of the mycelium to expose the fresh mycelium. The culture temperature is 18℃-20℃, and the relative humidity is maintained at 85%-95%.

[0073] (7) Cold stimulation: Place the mushroom bag in a 4℃~8℃ environment for 10~12 hours to improve the uniformity of the mushroom buds.

[0074] (8) Fruiting Management: After the buds emerge, the temperature should be controlled at 20℃~22℃, the relative humidity should be maintained at 90%~95%, and the CO2 concentration should be controlled at 3000ppm~10000ppm. The CO2 concentration should be controlled at 3000ppm~6000ppm on the first 3 days after the buds emerge; at around 10000ppm on the fourth day; and at around 5000ppm on the fifth day. Adequate ventilation should be ensured throughout the whole process.

[0075] The Pleurotus pulmonatus variety 'Shenxiu No. 2' of the present invention has high CO2 tolerance and can tolerate a CO2 concentration of 10,000 ppm.

[0076] (9) Harvesting: Harvest the whole flower and pack it in a fresh-keeping box.

[0077] (10) Management of multiple fruiting waves: After harvesting, clean the mushroom feet remaining in the bags. Rest for 12 to 13 days, spray the bags twice a day to keep them moist, and ventilate. Spray heavy water to replenish water before the next wave of cold stimulation. After 0.5 to 1 day, when the temperature is below 10°C, cold stimulation is performed for 8 to 10 hours. Continue to manage and harvest the fruiting according to steps (8) and (9). A total of 4 to 6 waves can be harvested.

[0078] In summary, the present invention discloses a high CO2 tolerance and one-time harvesting Pleurotus ostreatus variety and its application. The Pleurotus ostreatus variety has excellent traits, meets the diversified market needs, is suitable for factory cultivation, and has good application and promotion prospects. The pulmonary-shaped Pleurotus ostreatus variety 'Shenxiu No. 2' has high CO2 tolerance and can tolerate a high concentration of CO2 of 10,000 ppm during the bag opening and fruiting period, thereby promoting primordium differentiation, increasing the number of effective buds, increasing the width of the cap, and improving the conversion and absorption of the culture medium. It has the characteristics of early maturity, which is about 10 days shorter than that of the currently cultivated varieties, and has a short cultivation cycle. Tolerance to high concentrations of CO2 also enables the pulmonary-shaped Pleurotus ostreatus variety of the present invention to inhibit the growth of miscellaneous bacteria and reduce the risk of disease. The present invention is convenient to operate, saves labor and time, and has concentrated and uniform mushrooms, which can be harvested and packaged in one go. It also has high yield characteristics, with an average first-wave yield of 198.25 g per bag. The cap of the fruiting body of the pulmonary-shaped Pleurotus ostreatus variety 'Shenxiu No. 2' is light brown and slightly larger. The ISSR molecular marker combination of the pulmonary-shaped Pleurotus ostreatus variety 'Shenxiu No. 2' has the specificity and specificity to identify the pulmonary-shaped Pleurotus ostreatus variety 'Shenxiu No. 2'.

[0079] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A high CO2 tolerant and one-time harvesting Pleurotus pulmonarius variety, characterized by: The preservation number of the Pleurotus pulmonarius variety is GDMCC No: 65787; it was preserved in the Guangdong Provincial Microbiological Culture Collection Center on March 7, 2025.

2. Use of the high CO2 tolerance and one-time harvesting Pleurotus pulmonarius variety according to claim 1 in food production.

3. Use of the high CO2 tolerance and one-time harvesting Pleurotus pulmonarius variety according to claim 1 in Pleurotus pulmonarius breeding.

4. The factory cultivation method of the high CO2 tolerance and one-time harvesting Pleurotus pulmonarius variety according to claim 1, characterized in that: After the cultivation bag is sterilized by high-pressure steam and inoculated with the Pleurotus pulmonarius, it is moved into a culture room away from light for mycelium culture; when the mycelium fills the bag, it is subjected to after-ripening culture; scratching; cold stimulation culture; when young buds grow out, mushroom fruiting management is carried out; when mature, the whole mushroom is harvested and packaged at one time; 4 to 6 waves can be harvested.

5. The factory cultivation method of the high CO2 tolerant and one-time harvesting Pleurotus pulmonarius variety according to claim 4, characterized in that: The light-proof culture has a culture temperature of 23-25° C., a relative humidity of 65%-70%, and a culture time of 28-30 days.

6. The factory cultivation method of the high CO2 tolerant and one-time harvesting Pleurotus pulmonarius variety according to claim 4, characterized in that: The after-ripening culture has a culture temperature of 26 to 28° C. and a culture period of 8 to 10 days.

7. The factory cultivation method of the high CO2 tolerant and one-time harvesting Pleurotus pulmonarius variety according to claim 4, characterized in that: The scratching fungus has a culture temperature of 18-20° C. and a relative humidity of 85%-95%.

8. The factory cultivation method of the high CO2 tolerant and one-time harvesting Pleurotus pulmonarius variety according to claim 4, characterized in that: The cold stimulation culture has a cold stimulation temperature of 4-8° C. and a time of 10-12 hours. After the young buds grow out, the culture temperature is 20-22° C. and the relative humidity is 90%-95%.

9. The factory cultivation method of the high CO2 tolerant and one-time harvesting Pleurotus pulmonarius variety according to claim 4, characterized in that: The CO2 concentration during the mycelium culture period is controlled below 3500ppm; the CO2 concentration during the after-ripening culture period is controlled at 5500ppm~6000ppm; the CO2 concentration during the mushroom management period is controlled at 3000ppm~10000ppm.

10. The molecular identification method of Pleurotus pulmonarius varieties that are high CO2 tolerant and suitable for one-time harvesting according to claim 1, characterized in that: Molecular identification was performed using the following primers: ISSR 1: 5′-TGCACACACACACAC-3′; ISSR 2: 5′-GTGACACACACAC-3′; ISSR 3: 5′-GTGACGACTCTTCTCCTCTCT-3′; ISSR 4: 5′-GGATGCAACACACACACAC-3′; ISSR 5: 5′-CGTGTGTGTGTGTGT-3′; ISSR 6: 5′-AGTGTGTGTGTGTGT-3′.

Images