Novel black fungus strain and breeding method thereof

Abstract

According to the novel black fungus strain and the breeding method thereof, the novel black fungus strain is obtained by tissue isolation, wild domestication and systematic breeding of wild black fungus samples in a Tongling Dachuan forest farm, a Honggui forest farm and a Viltian forest farm, the novel black fungus strain is named as Auricularia auricula Haanjian No.1, the preservation number is CCTCC NO: M20241946, and the preservation number is CCTCC NO: M20241946. The collection date is September 9, 2024, and the collection site is China Center for Type Culture Collection. The invention provides a novel black fungus strain and a breeding method thereof.

Description

Technical Field

[0001] The invention belongs to the technical field of black fungus strains, and in particular relates to a new black fungus strain and a breeding method thereof. Background Art

[0002] Edible fungi refer to large fungi with large fruiting bodies that can be eaten, commonly known as mushrooms. There are more than 350 known edible fungi in China, most of which belong to the basidiomycetes, including: shiitake mushrooms, straw mushrooms, mushrooms, black fungus, white fungus, hericium, bamboo fungus, pine mushrooms (Matsutake), mushrooms, red mushrooms, ganoderma, cordyceps, truffles, white mushrooms and boletus; a few belong to the ascomycetes, including: morels, saddle mushrooms, truffles, etc.

[0003] Among them, black fungus, also known as black vegetable, fungus, and cloud fungus, belongs to the Auriculariaceae family, Auricularia genus. It is a precious colloid fungus that can be used as both medicine and food in my country. It is also a recognized health food in the world. The content of protein, vitamins and iron in dried black fungus is very high. Its protein contains a variety of amino acids, especially lysine and leucine. Summary of the invention

[0004] The invention provides a new black fungus strain and a breeding method thereof.

[0005] The technical solution adopted by the present invention is as follows: a new black fungus strain and a breeding method thereof, wherein the new black fungus strain is obtained by separating wild black fungus sample tissues from Wusongling Dachuan Forest Farm, Hutougou Forest Farm and Weihuling Forest Farm of Huangnihe Forestry Bureau in Dunhua City, Jilin Province, wild domestication and systematic breeding, and is named black fungus (Auricularia auricula) Hailan No. 1, with a preservation number of CCTCC NO: M20241946, a preservation date of September 9, 2024, a preservation location of China Center for Type Culture Collection, and a preservation unit address of Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province.

[0006] The biological characteristics of black fungus (Auricularia auricula) Hailan No. 1 are: single ear, high yield, light brown ear, multi-fold, medium-early maturing variety, growth period 80-92 days, white and dense mycelium, well-developed aerial mycelium, fresh ear light brown, multi-fold, single ear length 3.13-6.94cm, width 2.13-4.93cm, thickness 1.25-1.83mm, ear dry-wet ratio of 1:14, fresh fungus yield per 100kg dry material is 7.85kg.

[0007] A method for breeding a new black fungus strain comprises the following steps:

[0008] S1: Collect wild black fungus strains for initial screening;

[0009] S2: rescreen the four excellent strains initially screened;

[0010] S3: Conduct regional trials in randomized blocks;

[0011] S4: Conduct production trials in suitable planting areas;

[0012] S5: Physiological trait measurement;

[0013] S6: Statistics of natural incidence.

[0014] A method for growing a new black fungus strain comprises the following steps:

[0015] The spring ear is made into second-level seeds from October to December, and the third-level bags are made from January to March of the following year. The ears are put into the greenhouse to open and germinate in early April, and the ears are put into the field to open and germinate in late April to early May.

[0016] For autumn ear, the second-level seeds are made in January and February, and the third-level bags are made in April and May. They are put into the greenhouse to open and germinate in mid-July. They will open and germinate when the outside temperature rises to above 5-10℃.

[0017] Furthermore, the opening is a "three-sided" shaped opening with a diameter of 0.4 to 0.6 cm and a depth of 0.7 to 1 cm. The germination temperature is between 15 and 25° C. and the humidity is between 84% and 85%.

[0018] By adopting the method, the beneficial effects of the present invention are as follows: the variety is an excellent variety of black fungus with single ear, multiple folds, early maturity, high yield, uniform sprouting and good stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. They are used to explain the present invention together with the embodiments of the present invention and do not constitute a limitation of the present invention.

[0020] Figure 1 Antagonism diagram of the tested strains;

[0021] Figure 2 This is a bar graph showing the effect of different temperatures on the mycelium growth rate of "HaiLan No. 1 (HL111)";

[0022] Figure 3 This is a bar graph showing the effect of different pH values ​​on the mycelium growth rate of "Hai Lan No. 1 (HL111)";

[0023] Figure 4 This is a bar graph showing the effects of different carbon sources on the mycelial growth rate of “HaiLan No. 1 (HL111)”;

[0024] Figure 5This is a bar graph showing the effects of different nitrogen sources on the mycelial growth rate of “HaiLan No. 1 (HL111)”;

[0025] Figure 6 This is a bar chart showing the effects of different elements on the mycelial growth rate of "HaiLan No. 1 (HL111)";

[0026] Figure 7 This is a photo of the fruiting body of Heilan No. 1 (HL111);

[0027] Figure 8 These are photos of mycelium of Hailan No. 1 (HL111) and Yante No. 5 at 15℃;

[0028] Fig. 9 These are photos of mycelium of Hailan No. 1 (HL111) and Yante No. 5 at 20℃;

[0029] Fig.10 These are photos of mycelium of Hailan No. 1 (HL111) and Yante No. 5 at 25℃;

[0030] Fig.11 This is a photo of the antagonism between Heilan 1 (HL111) and Yante 5;

[0031] Fig.12 Photos showing the differences between Hailan No. 1 (HL111) and similar varieties (the left side shows Hailan No. 1 (HL111), and the right side shows similar varieties);

[0032] Fig.13 It is the case of small holes protruding from the ears;

[0033] Fig.14 To produce ear conditions;. DETAILED DESCRIPTION

[0034] A new black fungus strain and a breeding method thereof. The new black fungus strain is obtained by tissue separation, wild domestication and systematic breeding of wild black fungus samples from Wusongling Dachuan Forest Farm, Hutougou Forest Farm and Weihuling Forest Farm of Huangnihe Forestry Bureau in Dunhua City, Jilin Province, and is named (Auricularia auricula) Hailan No. 1, with a preservation number of CCTCC NO:M20241946, a preservation date of September 9, 2024, a preservation location of China Center for Type Culture Collection, and a preservation unit address of Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province.

[0035] The biological characteristics of black fungus (Auricularia auricula) Hailan No. 1 are: single ear, high yield, light brown ear, multi-fold, medium-early maturing variety, growth period 80-92 days, white and dense mycelium, well-developed aerial mycelium, fresh ear light brown, multi-fold, single ear length 3.13-6.94cm, width 2.13-4.93cm, thickness 1.25-1.83mm, ear dry-wet ratio of 1:14, fresh fungus yield per 100kg dry material is 7.85kg.

[0036] Secondary bacterial culture material formula: sawdust 77%, wheat bran 20%, bean flour 2%, gypsum 1%;

[0037] The formula of the third-level cultivation medium is: 81% sawdust, 15% wheat bran, 2% bean flour, 1.5% gypsum, 0.5% lime, and the water content is 62-65%.

[0038] A method for breeding a new black fungus strain comprises the following steps:

[0039] S1. Collect wild black fungus strains for preliminary screening:

[0040] Wild black fungus germplasm was collected from Wusongling Dachuan Forest Farm, Hutougou Forest Farm and Weihuling Forest Farm of Huangnihe Forestry Bureau in Dunhua City, Jilin Province, and tissue separation was performed. 18 strains were purified. After multiple mother strain and original strain culture and ear-out tests, the results showed that the mycelium growth rates of the 18 black fungus strains were 2.81-4.73 mm / d, and there were differences in the degree of mycelium development, colony morphology, uniformity and pigment production. Among them, the HL111 strain had the fastest mycelium growth rate, white mycelium, neat and dense colony edges, well-developed aerial mycelium, and pigment secretion (see Table 1 for details).

[0041] The results of the strain antagonism test showed that there were antagonistic lines between the strains, and obvious antagonistic reactions were produced. According to the results, it can be judged that the tested strains are somatically incompatible and all of them are strains with different physiological characteristics (see for details). Figure 1 ).

[0042] The results of the ear test of the tested strains showed that the growth period was between 75 and 107 days, with HL110 having the shortest growth period of 76 days and HL107 having the longest growth period (see Table 2 for details); the yield was 1.92-7.94g of dry ear per bag; among them, HL111 had the highest yield (see Table 2 for details).

[0043] Table 1 Mycelial growth

[0044]

[0045] Note: "+++", "++", "+" represent "developed", "medium", "underdeveloped" aerial hyphae respectively; "***", "**", "*" represent "dense", "medium", "sparse" colony morphology respectively; lowercase letters represent significant differences at the P < 0.05 level Duncan a,b ).

[0046] Table 2 Characteristics of tested strains

[0047]

[0048] Note: Lowercase letters indicate significant differences at the P < 0.05 level (Duncan analysis method), and the data are the average values ​​of 3 plots.

[0049] S2. Rescreen the 4 excellent strains initially screened:

[0050] The four selected excellent strains were subjected to repeated screening ear emergence test. The plot design adopted random block group, with three plots for each strain, and 100 bags of each strain were cultivated in each plot. The agronomic traits were compared through ear emergence test (details are shown in Table 3), and the excellent strain HL111 with multiple wrinkles, medium-early maturity and high yield was screened out. This strain was a wild strain collected from Hutougou Forest Farm of Huangnihe Forestry Bureau, Dunhua City, Jilin Province.

[0051] Table 3 Comparison of agronomic traits of superior strains

[0052]

[0053] Note: Lowercase letters indicate significant differences at the P < 0.05 level (Duncan analysis method), and the data are the average values ​​of 3 plots.

[0054] S3. Conduct regional trials in randomized blocks:

[0055] In Dunhua City, Wangqing County, Jiaohe City and other places in Jilin Province, with "Yante No. 5" as the control, the selected excellent strain "HL111" (Hai Lan No. 1) was carried out in multi-point regional tests. The experimental design of each test point uniformly adopted random block arrangement, 3 repetitions, 500 bags in each plot, and protection rows. The cultivation bag specifications were 17cm×36cm polyethylene folded corner bags, cooked material cultivation, and the management method was the same as conventional production. The average dry ear yield of the "Hai Lan No. 1" strain was 7.82kg / 100kg dry material, which was 8.46% higher than the control variety "Yante No. 5". The average growth period was 80-92 days. The ear pieces were light brown and wrinkled. The length of a single ear piece was 3.13-6.94cm, the width of the ear piece was 2.13-4.93cm, the thickness was 1.25-1.83mm, and the dry-wet ratio of the ear piece was 1:14. Multi-point tests for many years have shown high yield and medium-early maturity, proving that it has good stability (see Table 4 for details).

[0056] Table 4 Comparison of regional test results

[0057]

[0058] Note: Lowercase letters indicate significant differences at the P < 0.05 level (Duncan analysis method), and the data are the average values ​​of 3 plots.

[0059] S4. Carry out production trials in suitable planting areas:

[0060] Production trials were carried out in areas suitable for planting the experimental varieties, with "Yante 5" as the control variety. Each test site produced 1,500 bags, and the cultivation and management were the same as local production. The average dry ear yield of "HL111" (Hai Lan 1) was 7.85kg / 100kg dry material, 8.13% higher than the control variety "Yante 5". The average growth period was 80-92 days. The ear pieces were light brown with many wrinkles. The length of a single ear piece was 3.13-6.94cm, the width of the ear piece was 2.13-4.93cm, the thickness was 1.25-1.83mm, and the dry-wet ratio of the ear piece was 1:14. Multi-year multi-site trials have shown high yield and medium-early maturity (see Table 5 for details).

[0061] Table 5 Comparison of production test results

[0062]

[0063] Note: Lowercase letters indicate significant differences at the P < 0.05 level (Duncan analysis method), and the data are the average values ​​of 3 plots.

[0064] S5. Physiological characteristics measurement:

[0065] (1) High and low temperature tolerance test:

[0066] The 6mm diameter bacteria were inoculated into a 90mm PDA plate culture medium, and then placed in constant temperature incubators at 5℃, 10℃, 15℃, 20℃, 25℃, 30℃, and 35℃ respectively. They were cultured in the dark for 7 days, and the mycelium density and mycelium growth rate were recorded and measured. "HaiLan No. 1" can grow at 5℃ and 35℃, and has high and low temperature resistance.

[0067] (2) pH tolerance test:

[0068] The 6mm diameter bacteria were inoculated into 90mm PDA plate culture media with pH values ​​of 3, 4, 5, 6, 7, 8, 9, 10, 11, respectively. After inoculation, they were placed in a constant temperature incubator at 25℃ and cultured in the dark for 7 days. The mycelium density and mycelium growth rate were recorded and measured. "HaiLan No. 1" can grow at pH 5-11 and is resistant to acid and alkali.

[0069] (3) Optimal carbon source screening test:

[0070] The 6mm diameter bacteria were inoculated into 90mm plate culture media with lactose, sucrose, fructose, maltose and glucose as carbon sources respectively. After inoculation, they were placed in a constant temperature incubator at 25℃ and cultured in the dark for 7 days. The mycelium density and mycelium growth rate were recorded and measured. There was no significant difference in the mycelium growth rate of "HaiLan No. 1" in the culture media with sucrose, fructose and maltose as carbon sources.

[0071] (4) Optimal nitrogen source screening test:

[0072] The 6mm diameter bacterial strains were inoculated into 90mm plate culture media with corn flour, peptone, yeast extract powder, urea and ammonium sulfate as nitrogen sources. After inoculation, they were placed in a constant temperature incubator at 25℃ and cultured in the dark for 7 days. The mycelium density and mycelium growth rate were recorded and measured. The mycelium growth rate of "HaiLan No. 1" was the fastest in the culture medium with corn flour as nitrogen source.

[0073] (5) Optimal element screening test:

[0074] The 6mm diameter bacteria were inoculated into PDA basal medium, with natural pH, 0.15% KH2PO4, 0.1% MgSO4, 0.15% KH2PO4 + 0.1% MgSO4, 0.1% CaCO3, and 0.05% vitamin B1 as the trace elements for testing. After inoculation, they were placed in a 25℃ constant temperature incubator and cultured in the dark for 7 days. The mycelial growth rate of "Hai Lan No. 1 (HL111)" was the fastest in the medium with 0.1% MgSO4 and 0.15% KH2PO4 + 0.1% MgSO4 as elements.

[0075] S6. Statistics of natural incidence:

[0076] In the regional test of 1500 bags for each variety and the production test of 4500 bags for each variety, statistics were taken on fungi such as Trichoderma and Penicillium (see Table 6 for details). A small amount of contamination occurred in both "HaiLan No. 1" and "Yante No. 5", with a contamination rate of 1.54% for HaiLan No. 1 and 1.97% for Yante No. 5. The applied variety was slightly more resistant than the control variety.

[0077] Table 6 Field resistance survey of experimental varieties and control varieties

[0078]

[0079] A method for growing a new black fungus strain comprises the following steps:

[0080] The spring ear is made into second-level seeds from October to December, and the third-level bags are made from January to March of the following year. The ears are put into the greenhouse to open and germinate in early April, and the ears are put into the field to open and germinate in late April to early May.

[0081] For autumn ear, the second-level seeds are made in January and February, and the third-level bags are made in April and May. They are put into the greenhouse to open and germinate in mid-July. They will open and germinate when the outside temperature rises to above 5-10℃.

[0082] Among them, the opening is a "three-edged" shaped mouth with a diameter of 0.4-0.6cm and a depth of 0.7-1cm. The germination temperature is between 15-25℃, the humidity is between 84% and 85%, the young ear period of the strain is 5-7 days, and the humidity is between 84% and 85%. When the ear pieces grow to 2-3cm, stop watering for 1-2 days, and stop watering for 1-2 days when they grow to 4-5cm. Watering in the later period until harvesting. The best harvesting time is 6-7 minutes mature. The fruiting body is manifested by the contraction of the ear root, the expansion of the ear pieces, and the beginning of the ejection of spores on the ventral side. After the first harvest, stop watering for about 15-20 days to dry the bags. After the bags recover and harden, continue watering and the second batch of ears can be harvested.

[0083] Although the embodiments of the present invention have been shown and described, it is understood by those skilled in the art that various changes, modifications, substitutions and variations may be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents. In short, if those skilled in the art are inspired by them and design structural modes and embodiments similar to the technical solution without creativity without departing from the purpose of the present invention, they shall all fall within the scope of protection of the present invention.

Claims

1. A new strain of black fungus, characterized by: The new black fungus strain was obtained by isolation, wild domestication and systematic breeding of wild black fungus sample tissues from Wusongling Dachuan Forest Farm, Hutougou Forest Farm and Weihuling Forest Farm of Huangnihe Forestry Bureau in Dunhua City, Jilin Province, and was named Black Fungus (Auricularia auricula) Hailan No. 1, with a preservation number of CCTCC NO: M20241946, a preservation date of September 9, 2024, and a preservation location of China Center for Type Culture Collection.

2. A new black fungus strain according to claim 1, characterized in that: The biological characteristics of black fungus (Auricularia auricula) Hailan No. 1 are: single ear, high yield, light brown ear, multi-fold, medium-early maturing variety, growth period 80-92 days, white and dense mycelium, well-developed aerial mycelium, fresh ear light brown, multi-fold, single ear length 3.13-6.94cm, width 2.13-4.93cm, thickness 1.25-1.83mm, ear dry-wet ratio of 1:14, fresh fungus yield per 100kg dry material is 7.85kg.

3. The method for breeding a new black fungus strain according to claim 2, characterized in that: The following steps are involved: S1: Collect wild black fungus strains for initial screening; S2: rescreen the four excellent strains initially screened; S3: Conduct regional trials in randomized blocks; S4: Conduct production trials in suitable planting areas; S5: Physiological trait measurement; S6: Statistics of natural incidence.

4. The method for growing a new black fungus strain according to claim 3, characterized in that: The spring ear is made into second-level seeds from October to December, and the third-level bags are made from January to March of the following year. The ears are put into the greenhouse to open and germinate in early April, and the ears are put into the field to open and germinate in late April to early May. For autumn ear, the second-level seeds are made in January and February, and the third-level bags are made in April and May. They are put into the greenhouse to open and germinate in mid-July. They will open and germinate when the outside temperature rises to above 5-10℃.

5. The method for growing a new black fungus strain according to claim 4, characterized in that: The opening is a "triangular" shaped opening with a diameter of 0.4-0.6 cm and a depth of 0.7-1 cm. The germination temperature is between 15 and 25° C. and the humidity is between 84% and 85%.

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