Ganoderma sinensis new strain Guizhi No.8 and application thereof
By developing the new Zizhi strain Guizhi No. 8, the shortcomings of the existing varieties in selenium-enriched and utilizing wood culture medium were solved, and higher health care performance and commercial value were achieved.
Patent Information
- Application Number
- CN202510206928.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-25
- Publication Date
- 2025-06-13
AI Technical Summary
The existing purple zircon varieties have shortcomings in natural selenium-rich and the ability to utilize woody culture media, and it is difficult to find naturally selenium-rich varieties in germplasm resources through the prior art.
A new purple zhi strain, Guizhi No. 8, was developed. This strain was obtained through wild breeding and domestication. It has natural selenium-rich properties and has high yield laccase, cellulase and hemicellulase capabilities.
The No. 8 strain of Guizhi not only improves Zizhi's health care performance and its ability to utilize woody culture medium, but also provides Zizhi with better germplasm resources, significantly improving its commercial value.
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Abstract
Description
Technical Field
[0001] The present invention relates to the field of biotechnology, and particularly to a new strain of Ganoderma sinense, Guizhi No. 8, and its applications.
Background Art
[0002] Ganoderma sinense is a higher fungus that can be used both as medicine and food. Its main active ingredients include polysaccharides, triterpenoids, flavonoids, selenium, and sterols, and it has the effects of nourishing qi and calming the mind, relieving cough and asthma, alleviating fatigue, beautifying the skin, and delaying aging. It has varying degrees of curative effects on diseases such as chronic bronchitis, coronary heart disease, hepatitis, hyperlipidemia, neurasthenia, tumors, leukopenia, and dermatomyositis. The wild resources of Ganoderma sinense are scarce. In order to improve the artificial cultivation ability of Ganoderma sinense, it is necessary to obtain new strains of Ganoderma sinense from the source of the germplasm.
[0003] In the prior art, Ganoderma belongs to wood-growing edible fungi and has a strong ability to secrete laccase and cellulase, which helps to enhance the decomposition of lignin and cellulose. If these enzymes can be extracted and applied, it will greatly expand the application of wood-based culture media in the prior art. Moreover, in the prior art, we also found that the selenium content in the fruiting bodies of Ganoderma is relatively low. If we want to produce Ganoderma varieties rich in selenium, we need to add selenium elements to the culture medium additionally, and basically no naturally selenium-containing varieties have been found. If there are naturally selenium-containing varieties, it will greatly improve the health care ability of Ganoderma. Therefore, in order to strengthen the utilization of Ganoderma sinense germplasm resources and develop more excellent germplasm resources, it is necessary to continuously optimize the Ganoderma sinense germplasm resources to cultivate more commercially viable Ganoderma varieties.
Summary of the Invention
[0004] In view of the above, the purpose of the present invention is to provide a new strain of Ganoderma sinense: Guizhi No. 8. This variety was selected and domesticated from the wild by the applicant. Compared with other existing Ganoderma sinense varieties, this variety has a better appearance and thicker flesh. It is a naturally selenium-rich Ganoderma variety, and it has a higher ability to produce laccase, cellulase, and hemicellulase compared to other varieties. Its health care performance and the utilization ability of wood-based culture media are better than other varieties, and it can provide more excellent germplasm resources for Ganoderma sinense.
[0005] To achieve the above purpose, the present invention screened out a strain of Ganoderma sinense Guizhi No. 8, whose preservation number is CGMCC NO.41073. It belongs to edible fungi and belongs to the species Ganoderma sinense. The said Ganoderma is preserved in the China General Microbiological Culture Collection Center, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the preservation number of CGMCC NO.41073 and the preservation date of December 25, 2023.
[0006] The present invention also includes the application of the Ganoderma lucidum strain Guizhi No. 8 as a parent in cross-breeding.
[0007] The present invention also includes the spores obtained from the Ganoderma lucidum strain Guizhi No. 8.
[0008] The present invention also includes the mycelium obtained from the Ganoderma lucidum strain Guizhi No. 8.
[0009] The present invention also includes the fruiting body obtained from the Ganoderma lucidum strain Guizhi No. 8.
[0010] The present invention also includes the application of the fruiting body of the Ganoderma lucidum strain Guizhi No. 8 in food processing.
[0011] The present invention also includes the application of the mycelium of the Ganoderma lucidum strain Guizhi No. 8 in the extraction of laccase, cellulase and / or hemicellulase.
[0012] The present invention has the following beneficial effects:
[0013] 1. The Ganoderma lucidum strain Guizhi No. 8 of the present application was screened and domesticated by the research group from the wild environment. The test results show that: there are significant differences in mycelial growth and fruiting body appearance between this strain and existing similar varieties. Through the determination of inclusions, it is also found that: this strain is a Ganoderma lucidum strain naturally rich in selenium, and has a high ability to secrete laccase, cellulase and hemicellulase, which can effectively decompose lignin and cellulose in the nutrient bag, effectively improve the utilization ability of cellulose-based culture media, and is a rare Ganoderma lucidum variety rich in health care selenium. This variety can greatly enhance the commercial value of Ganoderma lucidum, and at the same time, it can also provide richer germplasm resources for Ganoderma lucidum.
Description of the Drawings
[0014] Figures 1 - 2 It is the result of the antagonistic experiment between Guizhi No. 8 and similar varieties, where Figure 1 is the comparison between Guizhi No. 8 and Wuzhi No. 2. In the figure: Guizhi No. 8 is above and Wuzhi No. 2 is below; Figure 2 is the comparison between Guizhi No. 8 and Heizhi (Jiangdu): Guizhi No. 8 is above and Heizhi (Jiangdu) is below.
[0015] Figure 3 is the comparison diagram of the mycelial bundle thickness between Guizhi No. 8 strain and Wuzhi No. 2.
[0016] Figure 4 is the comparison diagram of the colony density and morphology between Guizhi No. 8 strain and Wuzhi No. 2.
[0017] Figure 5 It is a comparison diagram of the colony pigments of Ganoderma lucidum strain Guizhi No. 8 and Wuzhi No. 2.
[0018] Figure 6 They are the upper and lower surfaces of the fruiting body cap of Ganoderma lucidum strain Guizhi No. 8.
[0019] Figure 7 They are the upper and lower surfaces of the fruiting body cap of the strain of the approximate variety Wuzhi No. 2.
[0020] Figure 8 It is a sectional view of the fruiting body of Ganoderma lucidum strain Guizhi No. 8.
[0021] Figure 9 It is a microscopic picture of the basidiospores of Ganoderma lucidum strain Guizhi No. 8.
Specific Embodiments
[0022] The present invention will be further described below in conjunction with the drawings, examples and tests.
[0023] Example 1:
[0024] In this example, a strain of Ganoderma lucidum strain Guizhi No. 8 was screened out, and its deposit number is CGMCC NO. 41073. It belongs to edible fungi and the species of Ganoderma lucidum. The said Ganoderma lucidum is deposited in the China General Microbiological Culture Collection Center, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, deposit number CGMCC NO. 41073, and the deposit date is December 25, 2023.
[0025] The above-mentioned Ganoderma lucidum strain Guizhi No. 8 was isolated, domesticated and cultivated by the applicant from the wild Ganoderma lucidum in Daming Mountain, Shanglin County, Nanning City. The antagonism diagram of this strain and other varieties of Ganoderma lucidum can be seen in Figure 1 and Figure 2 , among which, Figure 1 is the antagonism diagram of Guizhi No. 8 and Wuzhi No. 2, with Guizhi No. 8 on the top and Wuzhi No. 2 on the bottom; Figure 2 is the antagonism diagram of Guizhi No. 8 and Ganoderma atrum (Jiangdu), with Guizhi No. 8 on the top and Ganoderma atrum (Jiangdu) on the bottom. From the antagonism diagram, there is an obvious antagonistic line between Guizhi No. 8 and similar varieties, and it can be determined as different varieties.
[0026] The fruiting body morphology of Guizhi No. 8 can be seen in Figure 6 and Figure 8 : Among them Figure 6 are the upper and lower surfaces of the fruiting body cap of Ganoderma lucidum strain Guizhi No. 8, Figure 8Cross-sectional comparison diagrams of the fruiting bodies of Guizhi No. 8 and Wuzhi No. 2; as can be seen from the figures, the fruiting body cap forms earlier, the stipe is shorter, the shape of the stipe is flat cylindrical, the cap is thick, the size of the cap is small, the transverse diameter of the cap / the length of the stipe is medium, the thickness of the cap / the transverse diameter of the cap is large, the cap is fan-shaped, the color of the upper surface of the fruiting body is purple-black, the color of the lower surface is gray, there are radial longitudinal ridges on the upper surface of the cap, the annulus on the upper surface of the cap is medium, the edge of the cap is round and blunt, the flesh is clearly stratified, the color of the flesh is brown, the texture of the stipe is hard, and the texture of the cap is hard.
[0027] From Figure 6 and Figure 7 it can be seen that compared with the approximate variety Wuzhi No. 2, the fruiting body of Guizhi No. 8 strain is rounder than the control variety. Judging from the annular patterns, both varieties have annular patterns. However, the annular patterns of the new variety Guizhi No. 8 are more obvious and regular, and the appearance is more marketable.
[0028] Example 2:
[0029] This example is for the morphological identification of the Ganoderma lucidum strain Guizhi No. 8 screened in Example 1. Comparative identification test of fruiting body traits:
[0030] Cultivation and fruiting experiment: (1) Preparation of stock culture: Transfer the PDA slant culture to sawdust medium and culture at 25 °C for later use. (2) Preparation of fungus bags: Each bag is filled with 1 / 2 - 2 / 3 volume of miscellaneous wood medium, sterilized at 0.14 MPa - 0.15 MPa (temperature 126 °C - 127 °C) for 2.5 h and cooled for later use. (3) Inoculation: Transfer the stock culture to the fungus bags, and inoculate 10 bags for each number. After the mycelium fills the bags for 3 - 7 d, transfer them to the culture rack for ventilation management. The temperature during the mycelium growth period is set at 20 °C - 30 °C, and the temperature during the fruiting period is set at 24 °C - 26 °C. Manage the entire production process routinely, and the duration of the entire cultivation experiment is 5 months.
[0031] Comparison of fruiting body agronomic traits: Implement according to the standard of "Testing Guidelines for distinctness, uniformity and stability of plant varieties - Ganoderma lucidum" of the agricultural industry standard of the People's Republic of China. All data are calculated and the average value is used. See Table 1 for specific traits:
[0032] Table 1 Comparative control chart of morphological characteristics of Guizhi No. 8 and approximate variety Wuzhi No. 2
[0033]
[0034] Judging from the experimental records in Table 1, the primordium formation time of Guizhi No. 8 is earlier than that of the approximate variety Wuzhi No. 2, and the growth rate of the mycelium at 10 - 30 °C is significantly faster than that of the approximate variety Wuzhi No. 2.
[0035] The fruiting body morphology of Guizhi No. 8 is shown inFigure 6 and Figure 8 : Among them Figure 6 is the upper and lower surface diagrams of the fruiting body cap of Guizhi No. 8 strain, Figure 8 is the cross-sectional comparison diagram of the fruiting bodies of Guizhi No. 8 and Wuzhi No. 2; It can be seen from the figure that the fruiting body cap forms earlier, the stipe length is shorter, the stipe shape is flat cylindrical, the cap thickness is thick, the cap size is small, the cap transverse diameter / stipe length is medium, the cap thickness / cap transverse diameter is large, the cap is fan-shaped, the color of the upper surface of the fruiting body is purple-black, the color of the lower surface is gray, there are radial longitudinal ridges on the upper surface of the cap, the annulus on the upper surface of the cap is medium, the cap edge is round and blunt, the flesh is obviously layered, the flesh color is brown, the stipe texture is hard, the cap texture is hard, the spores are oval, and the spore length / width is large.
[0036] From Figure 6 and Figure 7 it can be seen that compared with the approximate variety Wuzhi No. 2, the fruiting body of Guizhi No. 8 strain is rounder than the control variety. From the annular patterns, both varieties have annular patterns. However, the annular patterns of the new variety Guizhi No. 8 are more obvious and regular, and the appearance is more marketable.
[0037] Example 3:
[0038] Analysis of the differences in the internal substances between the new strain Guizhi No. 8 and the approximate variety: Wuzhi No. 2
[0039] 1. Nutritional components:
[0040] Under the same conditions, Guizhi No. 8 and the approximate variety: Wuzhi No. 2 were cultivated to produce mushrooms. After fruiting, the fruiting bodies of the same batch with consistent maturity were collected, and the protein, polysaccharide and total amino acid contents of the Ganoderma sinense fruiting bodies were detected according to the relevant detection standards for nutritional components. The specific analysis results are shown in Table 2:
[0041] Table 2 Comparison of nutritional components between Guizhi No. 8 and its parental strains
[0042]
[0043] Note: Data in the same row with the same lowercase letter superscript or no superscript indicate no significant difference (P>0.05), and different lowercase letter superscripts indicate significant difference (P<0.05).
[0044] It can be seen from Table 2 that through detection and analysis, the total flavonoid, polysaccharide and selenium contents of the new strain Guizhi No. 8 are all significantly higher than those of the approximate variety Wuzhi No. 2 (P<0.05), and the total phenol and triterpene contents are all significantly lower than those of the approximate variety Wuzhi No. 2 (P<0.05). Most significantly, the Ganoderma sinense variety of this application is a natural selenium-containing strain, while other Ganoderma sinense varieties do not have this characteristic.
[0045] 2. Extracellular enzyme activity:
[0046] The applicant also conducted an assay on the extracellular enzyme activity of the mycelium of the new strain Guizhi No. 8 and similar varieties, as follows:
[0047] (1) Preparation of crude enzyme solution: After the Ganoderma lucidum mycelium filled the cultivation bags, 20 cultivation bags were taken from each generation of the strain. The upper 5 cm of the substrate was taken, freeze-dried, ground at low temperature through a 20-mesh sieve, and after mixing evenly, an appropriate amount of the substrate powder was taken. Distilled water or extraction solution was added at a mass-to-volume ratio of 1:10, and the supernatant obtained after soaking in a constant-temperature water bath and centrifugation was the crude enzyme solution.
[0048] (2) Enzyme activity assay: The ABTS method was used to assay the laccase activity: The amount of enzyme required to oxidize 1 μmoL of the substrate ABTS per minute per milligram of protein was defined as one enzyme activity unit; the 3,5-dinitrosalicylic acid (DNS) colorimetric method was used to assay the activities of cellulase, hemicellulase, and filter paper enzyme: The amount of enzyme required to catalyze the production of 1 mg of glucose per minute per milligram of protein was defined as one enzyme activity unit respectively.
[0049] Table 3 Results of the assay on the extracellular enzyme activity of the mycelium
[0050]
[0051] Note: The same lowercase letter or no letter as the superscript for the data in the same row indicates no significant difference (P > 0.05), and different lowercase letters as the superscript indicate a significant difference (P < 0.05).
[0052] As can be seen from Table 3, the activities of laccase, hemicellulase, and cellulase of Guizhi No. 8 are greater than those of the similar varieties, indicating that the new strain Guizhi No. 8 of this application has a better decomposition ability for lignin and cellulose.
[0053] Example 4:
[0054] The above Ganoderma sinense strain Guizhi No. 8 can be used as a parent to hybridize with other Ganoderma sinense strains of parents to obtain Ganoderma sinense germplasm resources with richer colors and more special morphologies.
[0055] In summary, the Ganoderma sinense strain Guizhi No. 8 of the present application was screened and domesticated by the applicant from the wild environment. It was verified and found that: the mycelial strands of this strain are finer, the mycelia are denser, the colony morphology is fluffier, the colony has pigments, the mycelial growth rate is slower, the primordium formation time is earlier, the fruiting body pileus formation time is earlier, the stipe length is shorter, the stipe shape is flat cylindrical, the pileus thickness is thick, the pileus size is small, the pileus transverse diameter / stipe length is medium, the pileus thickness / pileus transverse diameter is large, the pileus is fan-shaped, the upper surface color of the fruiting body is purple-black, the lower surface color is gray, there are radial longitudinal ridges on the upper surface of the pileus, the ring pattern on the upper surface of the pileus is medium, the pileus edge is round and blunt, the context is obviously layered, the context color is brown, the stipe texture is hard, the pileus texture is hard, the spores are oval, the spore length / width is large. In terms of the contained substances: the selenium content of the fruiting body is higher than that of the existing Ganoderma sinense varieties. In terms of enzyme activity: the laccase, hemicellulase and cellulase activities of the above strain are all greater than those of the existing Ganoderma sinense varieties. It shows that the new strain Guizhi No. 8 of the present application has a better decomposition ability for lignin and cellulose and is a Ganoderma sinense strain with excellent performance.
[0056] The above description is a detailed description of the preferred feasible embodiment of the present invention, but the embodiment is not used to limit the patent application scope of the present invention. Any equivalent changes or modifications made under the technical spirit disclosed by the present invention shall fall within the patent scope covered by the present invention.
Claims
1. Ganoderma lucidum strain Guizhi No. 8, with the preservation number of CGMCC NO.41073.
2. Use of the Ganoderma lucidum strain Guizhi No. 8 as claimed in claim 1 as a parent in hybrid breeding.
3. Spores obtained by culturing the Ganoderma lucidum strain Guizhi No. 8 as claimed in claim 1.
4. Mycelium obtained by culturing the Ganoderma lucidum strain Guizhi No. 8 as claimed in claim 1.
5. A fruiting body obtained by culturing the Ganoderma lucidum strain Guizhi No. 8 as claimed in claim 1.
6. Use of the fruiting body according to claim 5 in food processing.
7. Use of the mycelium according to claim 4 in extracting laccase, cellulase and / or hemicellulase.