Improved culture medium for agaricus bisporus mother culture and application thereof

Abstract

The application discloses an improved PDA culture medium for Agaricus bisporus mother culture and application and belongs to the technical field of edible mushroom cultivation. The improved PDA culture medium for Agaricus bisporus mother culture is based on a PDA culture medium and is added with Agaricus bisporus residue and peptone or beef extract. The culture medium provided by the application can significantly promote the growth of Agaricus bisporus mycelium, shortens the mother culture time from 23-25 days of the traditional method to about 13-15 days, greatly improves the strain production efficiency, and reduces the production cost and pollution risk.

Description

Technical Field

[0001] This invention belongs to the field of edible fungi cultivation technology, specifically relating to an improved PDA culture medium for button mushroom mother cultures and its application. Background Technology

[0002] Button mushrooms (Agaricus bisporus) are one of the most widely cultivated and consumed edible fungi worldwide. According to statistics from the China Edible Fungi Association, my country's total button mushroom production reached 1.4514 million tons in 2024, accounting for 3.31% of the country's total edible fungi production. In the past three years, button mushrooms have become one of the most profitable varieties in the edible fungi industry. Henan Province alone produced 140,450 tons of button mushrooms in 2024, a 21.44% increase compared to 2023. With the continuous expansion of production scale, the market demand for high-quality mushroom spawn is also constantly increasing.

[0003] The production of mother spawn (primary spawn) is a fundamental step in spawn propagation, and its growth rate and quality directly affect the production efficiency and final yield of subsequent primary and cultivars. Currently, the production of button mushroom mother spawn generally follows the national standard (GB 19171-2003), using potato dextrose agar (PDA) medium with the following formula: 200g potato (using extract), 20g glucose, 20g agar, and 1L water, with a natural pH. However, on this medium, the mycelium of button mushrooms grows slowly; under suitable temperature conditions (24℃±1℃), it typically takes 23 to 25 days for the mycelium to fully grow on a 9cm plate. This longer cultivation period not only increases production and time costs but also raises the probability of contamination by other microorganisms.

[0004] To shorten the mother culture cycle, those skilled in the art have attempted to improve PDA culture media. For example, invention patent (ZL202110643550.5) discloses a mother culture media containing Agaricus bisporus mycelium residue, with the following formula: 50-200g Agaricus bisporus mycelium residue, 200g potato, 20g glucose, 20g agar powder, and distilled water. This medium increases the mycelial growth rate to 3.77-4.75mm / d, requiring approximately 19 to 24 days for full growth on a 9cm plate. Although this is an improvement over traditional PDA culture media, the culture cycle is still relatively long, making it difficult to meet the industry's demand for high-efficiency production.

[0005] Therefore, providing a culture medium formulation that can further shorten the culture cycle of button mushroom mother culture and significantly promote mycelial growth is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0006] In view of this, the purpose of this invention is to provide an improved PDA culture medium for Agaricus bisporus mother culture and its application. The culture medium prepared by this invention can significantly improve the mycelial growth rate of Agaricus bisporus mother culture.

[0007] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a PDA-improved culture medium for button mushroom mother cultures, which consists of PDA culture medium and button mushroom residue.

[0008] Preferably, the amount of button mushroom residue added to each liter of PDA culture medium is 3-6g.

[0009] Preferably, the PDA culture medium for the button mushroom mother culture also includes peptone and / or beef extract.

[0010] Preferably, the amount of peptone added to each liter of PDA medium is 0.5-1g.

[0011] Preferably, the amount of beef extract added to each liter of PDA culture medium is 0.5-1g.

[0012] Preferably, the formula for 1L of PDA culture medium is: 200g potato, 20g glucose, 20g agar, and distilled water to a final volume of 1L.

[0013] This invention also provides the application of the above-mentioned PDA-modified culture medium for Agaricus bisporus mother culture in the rapid culture of Agaricus bisporus mother culture.

[0014] Preferably, the culture temperature is 24℃±1℃.

[0015] Preferably, the mycelial growth rate is 5.15-6.75 mm / d.

[0016] It contains at least the following beneficial technical effects: This invention achieves a synergistic effect by simultaneously adding specific amounts of Agaricus bisporus mycelium residue and peptone or beef extract to PDA culture medium. Experiments show that using the culture medium of this invention (PDA + 5g mycelium residue + 0.5g peptone), the mycelial growth rate of Agaricus bisporus can reach 6.75mm / d, which is much higher than the 3.87mm / d of traditional PDA culture medium and the 5.15mm / d of culture medium with only mycelium residue added.

[0017] The significantly increased mycelial growth rate directly shortens the mother culture cycle. Using the culture medium of this invention, inoculated onto a 9cm plate, it only takes about 13-15 days to reach full colony growth under suitable temperature, while traditional PDA medium requires 23-25 ​​days. This greatly improves the efficiency of mycelial production and reduces time and labor costs.

[0018] The shortened culture cycle reduces the time the strain is exposed to potential contaminants during the culture process, thereby effectively reducing the chance of contamination by other microorganisms and improving the success rate of strain production.

[0019] Button mushroom residue is a byproduct of mushroom production. It is widely available and inexpensive. Recycling it for use in the preparation of culture media not only reduces the cost of the culture media but also achieves resource recycling. Attached Figure Description

[0020] Figure 1 The mycelial growth of PDA cultured for 11 days; Figure 2 The mycelial growth of 5g of PDA + fungal residue cultured for 11 days; Figure 3 The mycelial growth after 11 days of culture with PDA + 5g of mycelial residue + 0.5g of peptone; Figure 4 The mycelial growth after 11 days of culture with PDA + 5g of mycelial residue + 1g of peptone; Figure 5 The mycelial growth of PDA + 5g of mushroom residue + 0.5g of beef extract cultured for 11 days; Figure 6 The mycelial growth of PDA + 5g of bacterial residue + 1g of beef extract cultured for 11 days. Detailed Implementation

[0021] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0022] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0023] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0024] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This application specification and embodiments are merely exemplary.

[0025] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0026] Unless otherwise specified, "room temperature" and "normal temperature" in this invention refer to 24±1℃.

[0027] Unless otherwise specified, all raw materials or instruments used in the following embodiments of the present invention are commercially available.

[0028] Example 1 Comparison of the preparation and effects of different culture medium formulations Test strain: Agaricus bisporus strain W192.

[0029] Basic culture medium (PDA) formulation: Peel 200g of potatoes, cut into chunks, add distilled water and boil for 20-30 minutes. Filter the extract through double-layered gauze. Add 20g of glucose and 20g of agar powder, and add distilled water to bring the volume to 1L. The pH should be natural. After dispensing, autoclave at 121℃ for 20 minutes.

[0030] Experimental formulation: Based on PDA medium, different amounts of Agaricus bisporus mycelium residue, peptone, beef extract, and yeast extract were added. Specific formulations and dosages are shown in Table 1. The Agaricus bisporus mycelium residue was added during the boiling of peeled potatoes. Other additives were added to the extract and thoroughly stirred to dissolve.

[0031] Preparation of button mushroom substrate: Collect the waste culture medium after harvesting button mushrooms, remove impurities, dry to constant weight, crush and sieve for later use.

[0032] Inoculation and Incubation: Pour approximately 20 mL of each of the above-mentioned culture medium formulations into 9 cm diameter petri dishes. After the culture medium has solidified, under aseptic conditions, use a 5 mm diameter punch to collect mycelial blocks from the edge of the pre-activated Agaricus bisporus W192 colony and inoculate them into the center of each plate. Set up 10 replicates for each formulation. Incubate the inoculated plates in a constant temperature incubator at 24℃±1℃ in the dark.

[0033] Data Measurement and Analysis: After the mycelium germinated and began to grow, the colony diameter was measured 11 days after inoculation using the cross-cross method. The growth rate (colony diameter) was calculated, and statistical methods were used to analyze the significant differences between different formulations. The results are shown in Table 1.

[0034] Table 1 Results analysis: Table 1 shows that adding 5g of Agaricus bisporus residue to PDA medium significantly promoted mycelial growth (from 3.87mm / d to 5.15mm / d). Adding appropriate amounts of peptone or beef extract to the residue further significantly promoted mycelial growth. The formula "PDA + 5g residue + 0.5g peptone" showed the most significant effect, with a mycelial growth rate of 6.75mm / d, significantly higher than other formulas. However, the mycelial growth rate of all formulas with added yeast extract was lower than that of the formulas with only residue, indicating that the yeast extract had an inhibitory effect in this combination.

[0035] Using this formula, the mycelial growth rate can reach 6.75 mm / d. Under 9 cm plate culture conditions, it only takes about 13-15 days to fully grow. Compared with the 23-25 ​​days of traditional PDA medium, the culture cycle is shortened by 76.9%, achieving unexpected technical results.

[0036] The above description is only a preferred embodiment of the present invention. It should be noted that those skilled in the art can make several improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A PDA-modified culture medium for Agaricus bisporus mother culture, characterized in that, The modified PDA culture medium for the button mushroom mother culture consists of PDA culture medium and button mushroom residue.

2. The PDA-modified culture medium for Agaricus bisporus mother culture according to claim 1, characterized in that, The amount of button mushroom residue added to each liter of PDA culture medium is 3-6g.

3. The improved PDA culture medium for Agaricus bisporus mother culture according to claim 1 or 2, characterized in that, The modified PDA culture medium for the mother mushroom of Agaricus bisporus also includes peptone or beef extract.

4. The PDA-modified culture medium for Agaricus bisporus mother culture according to claim 3, characterized in that, The amount of peptone added per liter of PDA medium is 0.5-1g.

5. The improved PDA culture medium for Agaricus bisporus mother culture according to claim 3, characterized in that, The amount of beef extract added to each liter of PDA culture medium is 0.5-1g.

6. The improved PDA culture medium for Agaricus bisporus mother culture according to claim 1, characterized in that, The formula for each liter of PDA culture medium is: 200g potato, 20g glucose, 20g agar, and distilled water to a final volume of 1 L.

7. The application of the PDA-modified culture medium for Agaricus bisporus mother culture as described in any one of claims 1-6 in the rapid culture of Agaricus bisporus mother culture.

8. The application according to claim 7, characterized in that, The culture temperature was 24℃±1℃.

9. The application according to claim 7, characterized in that, The mycelial growth rate is 5.15-6.75 mm / d.

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