High-activity sparassis crispa culture medium capable of slowing down strain degradation
By using polysaccharide carbon sources and citric acid chelated iron in the culture medium of *Hydrangea hydrangea*, the problem of strain degeneration of *Hydrangea hydrangea* was solved, mycelial vitality and yield were improved, and efficient cultivation and preservation of *Hydrangea hydrangea* were achieved.
Patent Information
- Application Number
- CN202511761818.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-27
- Publication Date
- 2026-01-13
AI Technical Summary
Hydrangea spore strains are prone to degeneration during cultivation and preservation, manifested as slower mycelial growth, reduced pigment secretion, and decreased yield, which affects the healthy and sustainable development of the edible fungi industry.
By using polysaccharide carbon sources such as glucomannan, xylan, and soybean oligosaccharides to replace monosaccharides in conventional culture media, and combining them with citric acid to chelate iron, the culture medium formula for *Hylocereus undatus* was optimized to enhance mycelial vigor and degradation capacity.
It significantly improves the activity of mycelial cellulose and hemicellulase, slows down strain degeneration, maintains high strain activity and stability, prevents pigment secretion, and increases yield.
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Figure CN121320104A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of preservation, propagation and preparation of edible fungus strains, and particularly relates to a high-vigor and strain-degradation-reducing Sparassis culture medium. BACKGROUND
[0002] In the cultivation, preservation and propagation of original and excellent edible fungus strains, due to internal genetic variation or external environmental factors, the excellent characteristics (such as high yield, high quality and strong stress resistance) of the strains are gradually weakened or lost and are inherited by the offspring. The decline and degradation of strain vigor are common and serious in the production of edible fungi. In the mycelial growth stage, the decline and degradation of strain vigor often appear as slow mycelial growth rate, sparse and weak mycelia, no longer thick, white and robust mycelia, easy to produce pigments, and the like, which result in the decline of the ability to decompose and utilize culture medium, abnormal fruiting bodies, and reduced yield, and the like, and seriously restrict the healthy and sustainable development of the entire edible fungus industry.
[0003] The direct manifestations of the degradation of Sparassis strain are slow growth of mycelia on the plate and secretion of pigments. Figure 1 Currently, in the production of Sparassis, the problems of weakened strain resistance, reduced yield and abnormal mushrooms caused by strain degradation often occur. Sparassis cultivation mainly uses coniferous wood chips such as pine and fir as the main raw material, and mainly utilizes cellulose and hemicellulose. Unlike broad-leaved trees, the hemicellulose of coniferous trees is mainly galactomannan, the main chain of which is composed of glucose and mannose, and the galactosyl group on the side chain is connected to the C6 position of the main chain mannose by an alpha-1,6 glycosidic bond. Therefore, there is an urgent need for an excellent culture medium formula that can effectively slow down the degradation of Sparassis strain and has strong cellulase and hemicellulase activity related to the degradation of mycelial substrate, so as to reduce the risk of strain degradation.
[0004] Currently, PDA is mainly used for daily preservation and transfer of Sparassis, and the influence of monosaccharides such as glucose, fructose and sucrose or starch substances on mycelial growth is mainly studied. SUMMARY
[0005] The purpose of the present application is to provide a high-vigor and strain-degradation-reducing Sparassis culture medium. According to the structural characteristics of the hemicellulose of coniferous trees, a polysaccharide similar in structure to the hemicellulose is selected as the carbon source, and a degradation-preventing culture medium with strong mycelial vigor and no pigment secretion is obtained by optimization, so as to slow down the problems of reduced strain vigor and easy degradation in daily production.
[0006] In order to achieve the above purpose, the technical scheme adopted by the present application is as follows:
[0007] A high-vigor and strain-degradation-reducing Sparassis culture medium comprises a basic culture medium and a polysaccharide carbon source.
[0008] Furthermore, the polysaccharide carbon source is one or more of glucomannan, xylan, soybean oligosaccharides, and galactan.
[0009] Furthermore, the amount of polysaccharide carbon source added is 10-30 g / L.
[0010] Furthermore, the preparation method of the basic culture medium is as follows: 50g to 100g of potatoes are extracted with boiling water and filtered to obtain potato extract. 2g of peptone, 20g of agar, 1g of potassium dihydrogen phosphate, and 1g of magnesium sulfate are added, and water is added to make up to 1L.
[0011] Furthermore, the *Hydrangea sylvestris* culture medium also includes chelated iron citrate, with the amount of chelated iron citrate added being 0.1-0.5 g / L.
[0012] The method for cultivating *Hydrangea macrophylla* using the above-mentioned culture medium is as follows:
[0013] 1) Preparation of Hydrangea sylvestris culture medium: Soak 50g-100g of potatoes in boiling water and filter to obtain potato extract. Add 10g-30g of polysaccharide carbon source, 2g of peptone, 20g of agar, 1g of potassium dihydrogen phosphate, and 1g of magnesium sulfate. Add water to make up to 1L and set the pH to natural.
[0014] 2) Sterilization: After dispensing the prepared *Hydrangea fuciformis* culture medium, autoclave at 121℃ for 20 minutes, and then cool to make a solid culture medium.
[0015] 3) Inoculation and culture: Inoculate the normal growing or degenerated Hydrangea strains onto the above-mentioned Hydrangea culture medium and culture them in a constant temperature and dark environment at 24℃.
[0016] This invention has the following advantages: The *Hydrangea hydrangea* culture medium of this invention uses polysaccharides as the main carbon source, replacing monosaccharides in conventional culture media. This significantly increases the expression levels of genes related to cellulose and hemicellulose degradation enzymes in *Hydrangea hydrangea* mycelia during cultivation, thereby enhancing strain vigor and slowing down strain degeneration. The formulation of this invention is simple, highly stable, and easy to implement, which is of great significance for maintaining the stability and high vigor of *Hydrangea hydrangea* strains. Attached Figure Description
[0017] Figure 1 Figure showing pigment secretion of *Hydrangea macrophylla* strains at different stages of degeneration.
[0018] Figure 2 The images show the Petri dishes of *Hydrangea hydrangea* strain grown on the culture medium of Example 1 (left image shows the front of the Petri dish, right image shows the back of the Petri dish).
[0019] Figure 3 The images show the Petri dishes of *Hydrangea hydrangea* strain grown on the culture medium of Example 2 (left image shows the front of the Petri dish, right image shows the back of the Petri dish).
[0020] Figure 4 The images show the Petri dishes of *Hydrangea spp.* grown on the culture medium of Example 3 (left image shows the front of the Petri dish, right image shows the back of the Petri dish).
[0021] Figure 5 The images show the Petri dishes of *Hydrangea hydrangea* strain grown on Comparative Example 1 medium (left image shows the front of the Petri dish, right image shows the back of the Petri dish). Detailed Implementation
[0022] The present invention will now be described in further detail with reference to specific embodiments.
[0023] In this invention, a normal-morphologically normal *Hydrangea hydrangea* strain with pigment secretion was used, preserved by the Institute of Edible Fungi, Fujian Academy of Agricultural Sciences.
[0024] Example 1
[0025] (1) Take 50g of potatoes and boil them in 1000mL of water for 30 minutes. Filter the solution through 6 layers of gauze to obtain potato extract.
[0026] (2) Weigh 20g xylan, 2g peptone, 20g agar, 1g potassium dihydrogen phosphate, 1g magnesium sulfate, and 0.2g citric acid chelated iron. Mix well and add potato extract. Add water to 1L and allow the pH to remain at its natural level to obtain the culture medium.
[0027] (3) After dispensing the prepared culture medium, autoclave it at 121℃ for 20 minutes, cool it and pour it into sterile petri dishes.
[0028] (4) Take a normal morphological test tube of *Hydrangea macrophylla*, cut the inoculum into 3mm×3mm inoculation blocks, pick up the inoculation blocks with an inoculation needle, place them in the center of the culture dish in step (3), and place them in a 24℃ constant temperature incubator for 25 days in the dark.
[0029] Example 2
[0030] (1) Take 50g of potatoes and boil them in 1000mL of water for 30 minutes. Filter the solution through 6 layers of gauze to obtain potato extract.
[0031] (2) Weigh 20g of glucomannan, 2g of peptone, 20g of agar, 1g of potassium dihydrogen phosphate, 1g of magnesium sulfate, and 0.2g of citric acid chelated iron. Mix well and add potato extract. Add water to 1L and set the pH to natural.
[0032] (3) After dispensing the culture medium prepared in step (2), autoclave at 121°C for 20 minutes, cool and pour into sterile culture dishes.
[0033] (4) Take a normal morphological test tube of *Hydrangea macrophylla*, cut the inoculum into 3mm×3mm inoculation blocks, pick up the inoculation blocks with an inoculation needle, place them in the center of the plate culture medium in step (3), and place them in a 24℃ constant temperature incubator for 25 days in the dark.
[0034] Example 3
[0035] (1) Take 50g of potatoes and boil them in 1000mL of water for 30 minutes. Filter the solution through 6 layers of gauze to obtain the extract.
[0036] (2) Weigh out 15g of glucomannan, 2g of peptone, 20g of agar, 1g of potassium dihydrogen phosphate, 1g of magnesium sulfate, and 0.2g of citric acid chelated iron. Mix well and add potato extract. Add water to 1L and set the pH to natural.
[0037] (3) After dispensing the culture medium prepared in step (2), autoclave at 121°C for 20 minutes, cool and pour into sterile culture dishes.
[0038] (4) Take the degenerate hydrangea plate with obvious pigment secretion around the original inoculation block, cut the inoculation block into a 3mm×3mm inoculation block, pick up the inoculation block with an inoculation needle, place it in the center of the plate culture medium in step (3), and place it in a constant temperature incubator at 24℃ for 25 days in the dark.
[0039] Comparative Example 1
[0040] Using the commonly used PDA culture medium in current production as a control, whose main carbon source is glucose, the preparation steps are as follows.
[0041] (1) Take 200g of potatoes and boil them in 1000mL of water for 30 minutes. Filter the solution through 6 layers of gauze to obtain the extract.
[0042] (2) Weigh 20g of glucose, 2g of peptone, 20g of agar, 1g of potassium dihydrogen phosphate and 1g of magnesium sulfate, mix well and add potato extract, add water to 1L, and set the pH to natural.
[0043] (3) After dispensing the culture medium prepared in step (2), autoclave at 121°C for 20 minutes, cool and pour into sterile culture dishes.
[0044] (4) Take a normal morphological test tube of *Hydrangea macrophylla*, cut the inoculum into 3mm×3mm inoculation blocks, pick up the inoculation blocks with an inoculation needle, place them in the center of the plate culture medium in step (3), and place them in a 24℃ constant temperature incubator for dark culture for 25 days.
[0045] Figure 2 The image shows the results of the *Hydrangea spp.* strain growing on the culture medium of Example 1. The mycelium grows vigorously and is highly active, with the central mycelium agglomerating to form primordial protrusions.
[0046] Figure 3 The results shown are those of the *Hydrangea hydrangea* strain growing on the culture medium of Example 2, which are similar to the growth morphology of Example 1.
[0047] Figure 4The pigmentation phenomenon was greatly alleviated when the degenerated *Hydrangea hydrangea* strain was cultured in the medium of Example 3.
[0048] Figure 5 The results shown are those of the *Hydrangea spp.* strain growing on Comparative Example 1 medium. Compared to Example 1 and Example 2, the *Hydrangea spp.* strain inoculated on conventional PDA medium showed signs of degeneration with slight pigment secretion around the inoculation site.
[0049] Table 1 shows the relative expression levels of genes related to cellulose and hemicellulose degradation in the differentially expressed genes of Examples 1 and 2, with Comparative Example 1 as the control. The results in Table 1 show that, compared to the commonly used PDA medium, the culture media of Example 1 and Example 2 showed significant differences in the expression of genes related to cellulose degradation, such as glucanase and glucosidase, as well as genes related to hemicellulose degradation, such as mannanase, mannosidase, galactosidase, and xylosidase. These genes were significantly upregulated, thus indicating stronger strain vigor and resistance to degradation.
[0050] Table 1. Differences in the expression of cellulose / hemicellulose degradation-related genes in different carbon source culture media.
[0051]
Claims
1. A culture medium for *Hydrangea hydrangea* with high activity and reduced strain degeneration, characterized in that, It includes a basal culture medium and a polysaccharide carbon source.
2. The *Hydrangea macrophylla* culture medium with high activity and reduced strain degeneration according to claim 1, characterized in that, The polysaccharide carbon source is one or more of glucomannan, xylan, soybean oligosaccharides, and galactan.
3. The *Hydrangea hydrangea* culture medium with high activity and reduced strain degeneration according to claim 1, characterized in that, The amount of polysaccharide carbon source added is 10-30 g / L.
4. The *Hydrangea macrophylla* culture medium with high activity and reduced strain degeneration according to claim 1, characterized in that, The preparation method of the basic culture medium is as follows: 50g~100g of potatoes are extracted with boiling water and filtered to obtain potato extract. 2g of peptone, 20g of agar, 1g of potassium dihydrogen phosphate, and 1g of magnesium sulfate are added, and water is added to make up to 1L.
5. The *Hydrangea hygroscopica* culture medium with high activity and reduced strain degeneration according to claim 1, characterized in that, The *Hydrangea sylvestris* culture medium also includes chelated iron in citrate.
6. The *Hydrangea hydrangea* culture medium with high activity and reduced strain degeneration according to claim 5, characterized in that, The amount of citric acid chelated iron added is 0.1-0.5 g / L.