Exogenous cross protective agent and application thereof in ganoderma lucidum cultivation

Abstract

The invention relates to the technical field of agriculture, and discloses an exogenous cross protective agent and application thereof in ganoderma lucidum cultivation. By applying the exogenous cross protective agent, the autotoxicity effect in the ganoderma lucidum cultivation process is effectively relieved, the biological yield of ganoderma lucidum is remarkably increased, meanwhile, the pollution rate is reduced, and the stress resistance is enhanced. According to the present invention, the applicability of the ganoderma lucidum as the deep processing raw material is significantly improved, the development of the related large health products is promoted so as to substantially improve the economic value of the ganoderma lucidum, and the exogenous cross-protective agent has advantages of high cost benefit, simple operation, easy transportation and storage and the like, such that the significant promotion potential and the significant market prospect are provided. In addition, the method is not only suitable for cultivation of ganoderma lucidum, but also plays an important guiding role in cultivation of other types of ganoderma lucidum and other edible fungi.

Description

Technical Field

[0001] The present invention relates to the technical field of agriculture, and in particular, to an exogenous cross-protectant and its application in the cultivation of Ganoderma lucidum Background Art

[0002] Under specific stress conditions suffered by microorganisms, their tolerance to other types of stress such as acidity, heat, and salt is enhanced, and this phenomenon is called the cross-protection effect. The research on this effect has widely involved multiple disciplinary fields such as animal breeding, plant cultivation, clinical medicine, environmental protection, and food fermentation. With the continuous progress of research techniques, the research on the cross-protection mechanism of microorganisms has also been gradually deepened, and many key genes and proteins have been identified, aiming to enhance the stress tolerance of microorganisms and reduce or alleviate the autotoxic feedback inhibition effect.

[0003] One of the strategies to enhance the tolerance of microorganisms using the cross-protection mechanism is to apply exogenous protectants, and this method has significant development potential in improving the production efficiency of microorganisms. The addition of exogenous protectants can enhance the tolerance of strains. Bioactive protectants play a key role in microorganisms' response to various stresses. Research indicates that under suitable stress conditions, various active substances in microorganisms such as glutathione, trehalose, glycerol, and amino acids will accumulate, showing potential protective functions. Therefore, applying exogenous protectants has become an effective strategy to improve the tolerance of microorganisms under stress conditions.

[0004] Among the currently artificially cultivated edible fungi, the autotoxic feedback inhibition phenomenon of Ganoderma lucidum is particularly significant. Based on the results of previous metabolomics research on Ganoderma lucidum, organic acids and their derivatives of Ganoderma lucidum constitute the main part of the differential metabolites. In two Ganoderma lucidum varieties, the contents of most differential organic acids have increased, which is consistent with the decrease in the pH value of the cultivation substrate during the cultivation process. This may provide a reasonable explanation for the fact that the remaining nutrients in the substrate cannot be used again to grow Ganoderma lucidum with qualified phenotypes. The mycelium secretes organic acids into the substrate, changing the environmental state. Organic acids have a negative regulatory effect on the growth of fruiting bodies by participating in metabolic pathways (such as purine metabolism regulation) and metabolism in different environments or other pathways, thus forming autotoxic feedback inhibition. Therefore, there is an urgent need for a method that can effectively alleviate the autotoxic feedback inhibition phenomenon of Ganoderma lucidum while improving its tolerance. Summary of the Invention

[0005] Based on the metabolomics analysis of extracellular metabolites during the previous cultivation process of Ganoderma lucidum, the applicant of the present invention developed an exogenous cross-protectant for Ganoderma lucidum cultivation, aiming to reduce the mushrooming obstacles of Ganoderma lucidum, improve its biological efficiency, guide the direction and technology of innovative breeding, help solve the common problems in the edible mushroom industry, promote the reuse of mushroom bran as a cultivation medium for homologous edible mushrooms, alleviate the crisis of edible mushroom cultivation media, develop high-value utilization technologies of mushroom bran, and lay a foundation for realizing efficient cultivation of edible mushrooms. In view of this, the present invention proposes an exogenous cross-protectant and its application in Ganoderma lucidum cultivation, aiming to provide a solution that can effectively alleviate the autotoxic feedback inhibition phenomenon of Ganoderma lucidum and improve its tolerance.

[0006] The present invention proposes a preparation method of an exogenous cross-protectant for Ganoderma lucidum cultivation, including:

[0007] Mixing Pueraria lobata extract, Perilla seed extract, vitamin H, vitamin B9 and vitamin B12 to obtain the exogenous cross-protectant for Ganoderma lucidum cultivation;

[0008] Among them, the weight ratio of the Pueraria lobata extract, Perilla seed extract, vitamin H, vitamin B9 and vitamin B12 is 58 - 62:27 - 30:7 - 10:0.8 - 3:0.2 - 2.

[0009] Preferably, the preparation method of the Pueraria lobata extract includes:

[0010] Crushing Pueraria lobata, then performing water extraction and filtration in sequence, concentrating the combined filtrate, and finally preparing the Pueraria lobata extract by spray drying.

[0011] Preferably, the preparation method of the Perilla seed extract includes:

[0012] Crushing Perilla seeds, extracting oil by supercritical extraction, then performing water extraction on the Perilla seed powder after oil extraction, filtering after extraction, concentrating the combined filtrate, and finally preparing the Perilla seed extract by spray drying.

[0013] The present invention also proposes an exogenous cross-protectant for Ganoderma lucidum cultivation prepared according to the above preparation method.

[0014] The present invention also proposes an application of the above exogenous cross-protectant for Ganoderma lucidum cultivation, and the application is A1 or A2;

[0015] A1 is the application of the exogenous cross-protectant for Ganoderma lucidum cultivation in Ganoderma lucidum cultivation;

[0016] A2 is the application of the exogenous cross-protectant for Ganoderma lucidum cultivation in the preparation of Ganoderma lucidum cultivation products.

[0017] The present invention also provides a fertilizer, which comprises the exogenous cross-protectant for Ganoderma lucidum cultivation as described above.

[0018] The present invention also provides an application of the above fertilizer in Ganoderma lucidum cultivation.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0020] The present invention discloses an exogenous cross-protectant for Ganoderma lucidum cultivation, and its preparation and application methods. This protectant is based on the physiological growth characteristics of Ganoderma lucidum, integrating agricultural product processing, microbiology, biochemistry, nutrition, and previous research results, and is innovatively prepared through scientific and reasonable processing techniques. By applying the exogenous cross-protectant prepared by the present invention, the phenomenon of autotoxic feedback inhibition during Ganoderma lucidum cultivation is significantly reduced, thereby effectively improving the biological efficiency of Ganoderma lucidum, reducing the pollution rate, increasing the yield, and enhancing its stress resistance. The application of the present invention makes Ganoderma lucidum more suitable as a raw material for intensive processing, promotes the development of related big health products, and greatly enhances the economic value of Ganoderma lucidum. The exogenous cross-protectant provided by the present invention has low cost, is easy to use, and the powder form is convenient for transportation and storage, so it has significant promotion value and market potential. The invention is widely applicable to the cultivation process of Ganoderma lucidum. In addition, the present invention also has important guiding significance for the cultivation of other Ganoderma species and other edible fungi. Detailed Embodiments

[0021] The exemplary embodiments of the present disclosure will be described in more detail below. Although the exemplary embodiments of the present disclosure are shown below, it should be understood that the present disclosure can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided so that the present disclosure can be more thoroughly understood and the scope of the present disclosure can be fully conveyed to those skilled in the art. It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the embodiments.

[0022] Example 1

[0023] The composition components of the exogenous cross-protectant for Ganoderma lucidum cultivation include: Pueraria lobata extract, Perilla seed extract, vitamin H, vitamin B9, and vitamin B12. The formula of this exogenous cross-protectant consists of the following components in parts by mass: Pueraria lobata extract (58 parts), Perilla seed extract (27 parts), vitamin H (10 parts), vitamin B9 (3 parts), and vitamin B12 (2 parts). The exogenous cross-protectant for Ganoderma lucidum cultivation and its preparation and application methods involved in the present invention include the following steps:

[0024] First, the kudzu root is crushed, followed by extraction, filtration. The filtrates are combined and concentrated, and finally, the kudzu root extract is prepared by spray drying. The parameters for water extraction are set as follows: solid-liquid ratio 1 g: 35 mL, extraction temperature 50 °C, extraction time 61 minutes, extraction times 5 times. After combining the extraction solutions and filtering, they are concentrated; the parameters for spray drying are: inlet air temperature 191 °C, sample injection rate 101 mL / min, induced draft fan frequency 51 Hz, centrifugal spray head rotation frequency 401 Hz;

[0025] Next, the perilla seeds are pulverized, and the oil is extracted using supercritical fluid extraction technology. The perilla seed powder after oil extraction is subjected to water extraction, and after filtration, the filtrates are combined and concentrated. Finally, the perilla seed extract is prepared by spray drying. The pulverization particle size of the perilla seeds is 125 - 150 mesh; the conditions for supercritical extraction are: extraction pressure 42 Mpa, extraction temperature 47 °C, separation kettle pressure 9.2 Mpa, the dosage of entrainer (anhydrous ethanol) is 13 mL·g -1 , extraction time 2.9 hours; the conditions for water extraction are: the raw material is the perilla seed powder after the above-mentioned supercritical extraction, solid-liquid ratio 1 g: 35 mL, extraction temperature 50 °C, extraction time 61 minutes, extraction times 5 times. After combining the extraction solutions and filtering, they are concentrated; the parameters for spray drying are: inlet air temperature 191 °C, sample injection rate 101 mL / min, induced draft fan frequency 51 Hz, centrifugal spray head rotation frequency 401 Hz;

[0026] The kudzu root extract and the perilla seed extract are added with vitamin H, vitamin B9, and vitamin B12 in the formula and mixed evenly.

[0027] Through the above steps, an exogenous cross-protectant for Ganoderma lucidum cultivation can be prepared.

[0028] The usage method of the exogenous cross-protectant for Ganoderma lucidum cultivation described in the present invention is as follows:

[0029] Dissolve the exogenous cross-protectant for Ganoderma lucidum cultivation in water. After complete dissolution, add it to the Ganoderma lucidum cultivation material.

[0030] Usage period: During the process of making Ganoderma lucidum fungus sticks, add it to the mixing water, and after complete dissolution, then carry out the mixing process.

[0031] The usage amount is: for every kilogram of cultivation material (dry weight), add 1 gram of the cross-protectant.

[0032] Under the same cultivation conditions, compared with the traditional method, the exogenous cross-protectant for Ganoderma lucidum cultivation prepared by the present invention has an average reduction of 94.28% in the contamination rate of Ganoderma lucidum mushroom sticks and an increase of 19.01% in biological efficiency. The results show that the exogenous cross-protectant for Ganoderma lucidum cultivation prepared by the present invention effectively reduces the autotoxic feedback inhibition obstacle in Ganoderma lucidum cultivation, can significantly improve the biological efficiency of Ganoderma lucidum, and reduce the contamination rate. The comparison of cultivating Ganoderma lucidum by this embodiment and the traditional method is shown in Table 1.

[0033] Table 1

[0034]

[0035] Note: Compared with the traditional method, only the exogenous cross-protectant for Ganoderma lucidum cultivation of the present invention is used, and other cultivation management conditions are the same; the biological efficiency is calculated based on the dry weight of the fruiting body.

[0036] Example 2

[0037] The composition components of the exogenous cross-protectant used for Ganoderma lucidum cultivation include: Pueraria lobata extract, Perilla seed extract, vitamin H, vitamin B9, and vitamin B12. The formula of this exogenous cross-protectant consists of the following components in parts by mass: Pueraria lobata extract (62 parts), Perilla seed extract (30 parts), vitamin H (7 parts), vitamin B9 (0.8 part), and vitamin B12 (0.2 part). The exogenous cross-protectant for Ganoderma lucidum cultivation and its preparation and application method involved in the present invention include the following steps:

[0038] First, the Pueraria lobata is crushed, then extracted and filtered. After combining the filtrates, it is concentrated, and finally, the Pueraria lobata extract is prepared by spray drying. The conditions for the water extraction process are: solid-liquid ratio 1g:45mL, extraction temperature 69°C, extraction time 80 minutes, extraction times 6 times. After combining the extraction solutions and filtering, it is concentrated; the conditions for the spray drying process are: inlet air temperature 195°C, sample injection speed 150mL / min, induced draft fan frequency 55Hz, and centrifugal spray head rotation speed frequency 450Hz;

[0039] Next, the Perilla seeds are pulverized, the oil is extracted using supercritical fluid extraction technology, and the Perilla seed powder after extracting the oil is subjected to water extraction. After extraction, it is filtered, the filtrates are combined and concentrated, and finally, the Perilla seed extract is prepared by spray drying. The pulverization particle size of the Perilla seeds is 150 mesh; the supercritical extraction conditions are: extraction pressure 43Mpa, extraction temperature 48°C, separation kettle pressure 10.2Mpa, and the dosage of entrainer (anhydrous ethanol) is 15mL·g -1, the extraction time is 3.5 hours; the water extraction conditions are as follows: the raw material is the perilla seed powder after the above-mentioned supercritical extraction, the material-to-water ratio is 1 g: 45 mL, the extraction temperature is 69 °C, the extraction time is 80 minutes, the number of extractions is 6 times, and the extraction solutions are combined and filtered and then concentrated; the conditions for the spray drying process are: the inlet air temperature is 195 °C, the sample injection speed is 150 mL / min, the frequency of the induced draft fan is 55 Hz, and the rotational speed frequency of the centrifugal spray head is 450 Hz;

[0040] Add vitamin H, vitamin B9, and vitamin B12 to the pueraria extract and perilla seed extract according to the formula and mix evenly.

[0041] Through the above steps, an exogenous cross-protectant for Ganoderma lucidum cultivation can be prepared.

[0042] The usage method of the exogenous cross-protectant for Ganoderma lucidum cultivation described in the present invention is as follows:

[0043] Dissolve the exogenous cross-protectant for Ganoderma lucidum cultivation in water, and after fully dissolving, add it to the Ganoderma lucidum cultivation material.

[0044] Usage period: During the process of making Ganoderma lucidum mushroom sticks, add it to the mixing water, fully dissolve it, and then carry out the mixing process.

[0045] The usage amount is: 50 grams of cross-protectant is added to each kilogram of cultivation material (dry weight).

[0046] Under the same cultivation conditions, compared with the traditional method, when using the exogenous cross-protectant for Ganoderma lucidum cultivation prepared by the present invention, the average pollution rate of Ganoderma lucidum mushroom sticks is reduced by 99.59%, and the biological efficiency is increased by 53.71%. The results show that the exogenous cross-protectant for Ganoderma lucidum cultivation prepared by the present invention effectively reduces the self-toxic feedback inhibition obstacle in Ganoderma lucidum cultivation, can significantly improve the biological efficiency of Ganoderma lucidum, and reduce the pollution rate. The comparison between cultivating Ganoderma lucidum by the method of the present invention and the traditional method is shown in Table 2.

[0047] Table 2

[0048]

[0049] Note: Compared with the traditional method, the present invention only uses the exogenous cross-protectant for Ganoderma lucidum cultivation of the present invention, and other cultivation management conditions are the same; the biological efficiency is calculated based on the dry weight of the fruiting body.

[0050] Example 3

[0051] The components of the exogenous cross-protectant used in Ganoderma lucidum cultivation include: Pueraria lobata extract, Perilla seed extract, biotin, folic acid, and cobalamin. The formula of the exogenous cross-protectant consists of the following components in parts by mass: Pueraria lobata extract (60 parts), Perilla seed extract (30 parts), biotin (8 parts), folic acid (1.0 part), cobalamin (1.0 part). The exogenous cross-protectant for Ganoderma lucidum cultivation and its preparation and application methods involved in the present invention include the following steps:

[0052] First, crush Pueraria lobata, then extract and filter it. After combining the filtrates, concentrate them, and finally obtain Pueraria lobata extract by spray drying. The conditions for the water extraction process are: solid-liquid ratio 1 g: 40 mL, extraction temperature 60 °C, extraction time 70 minutes, extraction times 5 times. Combine the extraction liquids, filter them, and then concentrate. The conditions for spray drying are: inlet air temperature 192 °C, sample injection speed 140 mL / min, induced draft fan frequency 54 Hz, centrifugal spray head rotation frequency 440 Hz;

[0053] Second, powder Perilla seeds, extract the oil using supercritical extraction technology, perform water extraction on the Perilla seed powder after extracting the oil, filter after extraction, combine the filtrates and concentrate, and finally obtain Perilla seed extract by spray drying. The crushing particle size of Perilla seeds is 140 mesh; the conditions for supercritical extraction are: extraction pressure 42 Mpa, extraction temperature 47 °C, separation kettle pressure 9.2 Mpa, the dosage of entrainer (anhydrous ethanol) is 14 mL·g -1 , extraction time is 3.3 hours; the conditions for water extraction are: the raw material is the Perilla seed powder after the above-mentioned supercritical extraction, solid-liquid ratio 1 g: 40 mL, extraction temperature 60 °C, extraction time 70 minutes, extraction times 5 times. Combine the extraction liquids, filter them, and then concentrate. The conditions for spray drying are: inlet air temperature 192 °C, sample injection speed 140 mL / min, induced draft fan frequency 54 Hz, centrifugal spray head rotation frequency 440 Hz;

[0054] Add biotin, folic acid, and cobalamin to the Pueraria lobata extract and Perilla seed extract according to the formula, and mix them evenly.

[0055] Through the above steps, the exogenous cross-protectant for Ganoderma lucidum cultivation can be prepared.

[0056] The usage method of the exogenous cross-protectant for Ganoderma lucidum cultivation described in the present invention is:

[0057] Dissolve the exogenous cross-protectant for Ganoderma lucidum cultivation in water. After fully dissolving, add it to the Ganoderma lucidum cultivation material. That's it.

[0058] Usage period: During the process of making Ganoderma lucidum spawn bags, add it to the mixing water. After fully dissolving, then carry out the mixing process.

[0059] Dosage: For every kilogram of cultivation material (dry weight), 20 grams of the cross-protectant can be added.

[0060] Under the same cultivation conditions, compared with the traditional method, when using the exogenous cross-protectant for Ganoderma lucidum cultivation prepared by the present invention, the contamination rate of Ganoderma lucidum spawn bags is on average reduced by 99.22%, and the biological efficiency is increased by 38.18%. The results show that the exogenous cross-protectant for Ganoderma lucidum cultivation prepared by the present invention effectively reduces the autotoxic feedback inhibition obstacle in Ganoderma lucidum cultivation, can significantly improve the biological efficiency of Ganoderma lucidum, and reduce the contamination rate. The comparison between cultivating Ganoderma lucidum by the present invention and the traditional method is shown in Table 3.

[0061] Table 3

[0062]

[0063]

[0064] Note: Compared with the traditional method, the present invention only uses the exogenous cross-protectant for Ganoderma lucidum cultivation of the present invention, and other cultivation management conditions are the same; the biological efficiency is calculated based on the dry weight of the fruiting body.

[0065] Finally, 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 them. Although the present invention has been described in detail with reference to the above embodiments, those of ordinary skill in the art should understand that: still can modify the specific implementation manners of the present invention or make equivalent replacements, and any modification or equivalent replacement without departing from the spirit and scope of the present invention shall be covered by the protection scope of the claims of the present invention.

Claims

1. A preparation method of an exogenous cross-protectant for Ganoderma lucidum cultivation, characterized in that, Comprising: Mixing Pueraria lobata extract, Perilla seed extract, biotin, folic acid and cobalamin to obtain the exogenous cross-protectant for Ganoderma lucidum cultivation; Among them, the weight ratio of the Pueraria lobata extract, Perilla seed extract, biotin, folic acid and cobalamin is 58 - 62:27 - 30:7 - 10:0.8 - 3:0.2 - 2.

2. The preparation method of the exogenous cross-protectant for Ganoderma lucidum cultivation according to claim 1, wherein, The preparation method of the Pueraria lobata extract includes: Crushing Pueraria lobata, followed by water extraction and filtration in sequence, concentrating the combined filtrate, and finally preparing the Pueraria lobata extract by spray drying.

3. The preparation method of the exogenous cross-protectant for Ganoderma lucidum cultivation according to claim 1, wherein, The preparation method of the Perilla seed extract includes: Crushing Perilla seeds, extracting oil by supercritical fluid extraction, then performing water extraction on the Perilla seed powder after oil extraction, filtering after extraction, concentrating the combined filtrate, and finally preparing the Perilla seed extract by spray drying.

4. An exogenous cross-protectant for Ganoderma lucidum cultivation prepared by the preparation method according to claims 1 - 3.

5. Use of an exogenous cross-protectant for Ganoderma lucidum cultivation according to claim 4, characterized in that, The application is A1 or A2; The A1 is the application of the exogenous cross-protectant for Ganoderma lucidum cultivation in Ganoderma lucidum cultivation; The A2 is the application of the exogenous cross-protectant for Ganoderma lucidum cultivation in the preparation of Ganoderma lucidum cultivation products.

6. A fertilizer, characterized in that, The fertilizer includes the exogenous cross-protectant for Ganoderma lucidum cultivation according to claim 4.

7. An application of the fertilizer according to claim 6 in Ganoderma lucidum cultivation.

Images