Lentinus edodes cultivation material containing longan shells as well as preparation method and application of lentinus edodes cultivation material

Abstract

The invention discloses a lentinus edodes cultivation material containing longan shells as well as a preparation method and application of the lentinus edodes cultivation material. The formula of the mushroom cultivation material containing the longan shells comprises sawdust, the longan shells, bran, gypsum and lime. The beneficial effects of part of longan shells instead of wood chips on mushroom hypha growth, nutritional ingredients and flavor substances provide experimental basis for the application of the longan shells in the technical field of edible mushroom cultivation in the future, and the method is also the biggest innovation point of the invention.

Description

Technical Field

[0001] The invention relates to the field of recycling technology of longan shells and the field of edible fungus cultivation technology, and in particular to a mushroom cultivation material containing longan shells, and a preparation method and application thereof. Background Art

[0002] Shiitake mushrooms belong to the phylum Basidiomycota, class Agaricomycetes, order Agaricales, family Omphalotaceae, and genus Lentinula. They are a famous traditional edible fungus in my country with a history of nearly a thousand years of artificial cultivation. Shiitake mushrooms are tender and delicate, with a unique flavor and delicious taste. They are rich in nutrient active ingredients such as shiitake polysaccharides, sulfur-containing compounds (such as shiitake essence), terpenes, alkaloids, amino acids, trace elements, vitamins, etc. They are both medicinal and edible, and are deeply loved by consumers and well-known both at home and abroad. Modern active efficacy research shows that the active ingredients in shiitake mushrooms have significant effects in regulating immune function, inhibiting the growth of cancer cells, and improving various metabolic disorder-related diseases, showing huge high nutritional, medicinal and market value. According to the data of the China Edible Fungi Association, the output of fresh shiitake mushrooms in 2023 was 13.534 million tons, accounting for 32.05% of the total output of fresh edible fungi, making it the edible fungus with the highest output in my country. With the gradual expansion of the scale of shiitake mushroom production, traditional cultivation substrates such as sawdust are becoming increasingly scarce and their costs are increasing. Therefore, screening new substrates suitable for shiitake mushroom cultivation is an urgent task to be solved. Suitable new substrates not only reduce the excessive dependence of traditional shiitake mushroom cultivation on broadleaf sawdust and promote the healthy development of the shiitake mushroom cultivation industry, but also enhance the social, economic and ecological benefits of shiitake mushrooms.

[0003] Longan (Dimocarpus longan L.) is one of the most popular subtropical fruits in China. It has important economic value, unique taste, rich in active nutrients such as polysaccharides, polyphenols, and flavonoids, and has multiple effects such as anti-aging, lowering blood lipids, and antibacterial. According to the results of many studies, it was found that the external application of longan shell decoction can prevent skin allergies. It contains polysaccharides, flavonoids, quercetin, yellow pigment, proanthocyanidins and suberin, and shows great potential in related industries such as antibacterial and natural dyes. In the process of eating and processing longan, longan shells and longan cores, which account for about 1 / 3 of the fresh weight of longan, are discarded as by-products; with the rapid development of the longan processing production chain, the amount of these two wastes has increased year by year, causing certain pressure on the nearby environment and causing waste of resources.

[0004] Applying longan shell, a by-product of longan processing, to the shiitake mushroom cultivation-related industrial chain can not only reduce the pressure of longan shell on the environment and the waste of resources to a certain extent, but also alleviate the excessive dependence of shiitake mushroom production on forest resources. Summary of the invention

[0005] The purpose of the present invention is to provide a formula of shiitake mushroom cultivation material containing longan shells, a preparation method and application thereof, so as to improve the quality of shiitake mushrooms, provide a new direction for the recycling of longan shells, reduce the pressure on the environment, realize the green and efficient utilization of agricultural wastes, and improve the economic and ecological benefits.

[0006] The first object of the present invention is to provide a formula of mushroom cultivation material containing longan shell, comprising sawdust, longan shell, bran, gypsum and lime.

[0007] Preferably, the longan shell accounts for 15-60% by mass, more preferably 15-45%, and even more preferably 15-30%.

[0008] Preferably, the mixture comprises, by mass fraction, 50-65% of sawdust, 15-30% of longan shells, 18.8% of bran, 1% of gypsum and 0.2% of quicklime.

[0009] Preferably, by mass fraction, it comprises 65% sawdust, 15% longan shell, 18.8% bran, 1% gypsum and 0.2% quicklime. The second object of the present invention is to provide a method for preparing a mushroom cultivation material formula containing longan shell, comprising the following steps:

[0010] (1) drying longan shells in an oven at 50° C. to 60° C. and crushing them in a crusher;

[0011] (2) Mix with other planting materials.

[0012] The third object of the present invention is to provide application of longan shell in mushroom cultivation.

[0013] According to some embodiments of the present invention, it was found that longan shells have an effect on the growth of shiitake mushroom mycelium and the active ingredients and volatile components of fruiting bodies, and a low proportion of longan shells promotes the growth of shiitake mushroom mycelium and increases the content of crude protein, umami amino acids, sweet amino acids, bitter amino acids, total amino acids and shiitake essence in shiitake mushroom fruiting bodies.

[0014] Therefore, preferably, longan shell is used as a cultivation material for cultivating shiitake mushrooms to promote the growth of shiitake mushroom mycelium and increase the content of crude protein, umami amino acids, sweet amino acids, bitter amino acids, total amino acids and shiitake essence in shiitake mushroom fruiting bodies.

[0015] Further preferred is the use of the above-mentioned mushroom cultivation material formula in promoting the growth of mushroom mycelium and increasing the contents of crude protein, crude fat, total amino acids, umami amino acids, sweet amino acids, and bitter amino acids in fruiting bodies, thereby improving the quality of mushroom fruiting bodies.

[0016] Further preferred is the use of the above-mentioned mushroom cultivation material formula in increasing the content of main volatile flavor substances in mushroom fruiting bodies. Preferably, the volatile flavor substances are 1,2,3,5,6-pentathiacycloheptane (mushroom essence), 1,2,4,6-tetrathiacycloheptane, 1,2,4-trithiacyclopentane, carbonyl sulfide and methyl mercaptan.

[0017] Longan shell is a byproduct of longan processing, containing polysaccharides, flavonoids, quercetin, yellow pigment, proanthocyanidins and suberinone, and has great potential in related industries such as antibacterial and natural dyes. Applying longan shell to the industrial chain related to mushroom cultivation can not only reduce the pressure of longan shell on the environment and the waste of resources to a certain extent, but also reduce the excessive dependence of mushroom production on forest resources, and has high economic, ecological and social value.

[0018] According to some embodiments of the present invention, the beneficial effect of partially replacing sawdust with longan shells on the growth of shiitake mushroom mycelium and nutritional components and flavor substances provides an experimental basis for its future application in the field of edible fungus cultivation technology, which is also the biggest innovation of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The present invention relates to a group of Lentinus edodes fruiting bodies with different longan shell content formulas.

[0020] Figure 2 The effects of different longan shell contents on the growth rate (A), stipe diameter (B), stipe length (C), cap thickness (D), and cap diameter (E) of Lentinus edodes in the present invention. Control group: 80% sawdust; Formula 1: 65% sawdust and 15% longan shell; Formula 2: 50% sawdust and 30% longan shell; Formula 3: 35% sawdust and 45% longan shell; Formula 4: 20% sawdust and 60% longan shell. Note: Compared with the control group, *P<0.05 (statistically significant difference), **P<0.01 (significant statistical difference).

[0021] Figure 3 The effects of different longan shell contents on the contents of crude protein (A), crude fat (B), total sugar (C), total amino acids (D), umami amino acids (E), sweet amino acids (F), and bitter amino acids (G) in shiitake mushroom fruiting bodies in the present invention. Control group: 80% sawdust; Formula 1: 65% sawdust and 15% longan shell; Formula 2: 50% sawdust and 30% longan shell; Formula 3: 35% sawdust and 45% longan shell; Formula 4: 20% sawdust and 60% longan shell. Note: Compared with the control group, *P<0.05 (statistically significant difference), **P<0.01 (significant statistical difference), ***P<0.001 (extremely significant statistical difference).

[0022] Figure 4The effect of different longan shell contents on the contents of several different volatile aroma substances in the present invention. Control group: 80% sawdust; Formula 1: 65% sawdust and 15% longan shell; Formula 2: 50% sawdust and 30% longan shell; Formula 3: 35% sawdust and 45% longan shell; Formula 4: 20% sawdust and 60% longan shell. Note: dotted line, change multiple >2 or <0.5, P <0.05, importance value of variable to model >1. DETAILED DESCRIPTION

[0023] The present invention will be described in detail below in conjunction with the examples. The methods described in the examples of the present invention are all conventional methods unless otherwise specified.

[0024] Example 1

[0025] A method for cultivating shiitake mushrooms using longan shells comprises the following steps:

[0026] Preparation of mushroom cultivation material: Take 30kg of longan shells, dry them in an oven at 50℃~60℃, and crush them in a grinder (about 1cm 2 ), mixed with a certain proportion of sawdust and divided into a control group and 4 formulation groups:

[0027] Control group, 15 kg sawdust (80%);

[0028] Formula 1: 12.1875kg (65%) sawdust + 2.8125kg (15%) longan shell;

[0029] Formula 2: 9.375kg (50%) sawdust + 5.625kg (30%) longan shells;

[0030] Formula 3: 6.5625kg (35%) sawdust + 8.4375kg (45%) longan shells;

[0031] Recipe 4: 3.75kg (20%) sawdust + 11.25kg (60%) longan shells.

[0032] After mixing, they are pre-wetted in water and then mixed with auxiliary materials (3.025kg bran, 0.1875kg gypsum and 0.0375kg lime) to a water content of about 50%, and packed into polyethylene bags (17cm×60cm), with 2.5kg of wet material for each mushroom stick and 15 bags for each group.

[0033] Sterilization: high temperature and high pressure sterilization, sterilization parameters: 105℃30min, 115℃30min, 121℃180min, 120min. Inoculation: The strain is Shiitake Mushroom Shenxiang 215 (provided by the Vegetable Institute of Wuhan Academy of Agricultural Sciences), each mushroom stick is inoculated with three inoculation points, and the inoculated mushroom sticks are bagged.

[0034] Cultivation: The temperature of the cultivation room is controlled at 22℃~23℃, and the center temperature of the mushroom stick is controlled at about 25℃. After the mycelium at the three inoculation points germinates and grows in contact, the bagging is removed and the growth rate of mycelium in different groups is measured.

[0035] Compared with the mycelial growth rate of the control group (0.48±0.05cm / d), the mycelial growth rates of Formula 1, Formula 2 and Formula 3 were faster, which were 0.61±0.09cm / d, 0.50±0.06cm / d and 0.57cm / d±0.08cm / d, respectively. The mycelial growth rate of Formula 4 was the slowest (0.45±0.06cm / d). There were significant differences between Formula 1 and Formula 3 and the control group (P<0.05, Figure 2 A), the above results show that a certain amount of longan shell can promote the growth of Lentinus edodes mycelium.

[0036] Color change: When the mushroom sticks have finished spawning and begin to form nodules, puncture the holes to a depth of about 6 cm in diameter, and induce color change with light stimulation, and the color change temperature is controlled at 24-26°C. Fruiting management: The temperature in the mushroom house is controlled at 15-18°C, and the humidity is 85%-90%. After the primordium is formed (the hyphae are twisted to form the primordium on the same day), the mushrooms are cut and fruited, and the fruiting bodies are picked in time (the fruiting bodies are harvested for the first time on the same day).

[0037] Analysis of fruiting body traits: Select regular shaped half-ripe fruiting bodies (about to open) to measure agronomic traits such as cap diameter, cap thickness, stipe diameter, and stipe length. The cap diameter is measured as the distance between the two farthest points on the cap edge, the cap thickness is the thickness at the junction of the cap and the stipe, the stipe diameter is the data value at the point where the stipe is uniform in thickness, and the stipe length is the distance between the top and bottom of the stipe. Each group measures 10 to 20 fruiting bodies.

[0038] according to Figure 2 From B and C, we can see that the average stipe diameters of formula 1, formula 3 and formula 4 are larger than the average stipe diameter of the control group (16.50±2.27cm), which are 17.94±1.88cm, 18.46±5.03cm and 18.04±2.83cm respectively; the average stipe lengths of the four formula groups are all larger than the average stipe lengths of the control group. However, there are no significant differences in the average diameter and average length of the stipes of the four formula groups compared with the control group. Figure 2D and E show that the average cap diameter of formula 1 (60.69±5.40cm) is greater than that of the control group (53.26±4.93cm), while there is no significant difference in the average cap diameters of all groups; the average cap thickness of formula 4 (9.17±2.24cm) is significantly less than that of the control group (13.61±1.57cm) (P<0.001). The above results show that a certain amount of longan shell can help the growth of the stipe and cap of shiitake mushroom fruiting bodies.

[0039] Example 2

[0040] The shiitake mushroom fruiting bodies of different groups in the test example 1 were placed in a 50°C oven for drying until the weight was constant and no longer changed, then put into a ziplock bag and stored at 4°C. The changes in the nutritional active components were determined, mainly including crude protein, crude fat, total sugar, and amino acids. The test regulations are as follows:

[0041] (1) Crude protein determination: The Kjeldahl method was used in accordance with GB / T 15673-2009 “Determination of crude protein content in edible fungi”. Each group contained three parallel tests.

[0042] (2) Crude fat determination: The crude fat content was determined by Soxhlet extraction according to GB / T 15674-2003 “Determination of crude fat content in edible fungi”. Each group contained three parallel tests.

[0043] (3) Total sugar determination: The determination was carried out in accordance with NY / T 1676-2008 “Determination of crude polysaccharide content in edible fungi”. Each group contained three parallel tests.

[0044] (4) Amino acid determination: Liquid chromatography-mass spectrometry was used in accordance with T / NAIA 0206-2023 “Determination of free amino acids in edible fungi”. Each group contained three parallel tests.

[0045] The results are as follows Figure 3 As shown in A, the crude protein content in the fruiting bodies of Formula 1, Formula 2 and Formula 3 was significantly higher than that of the control group (193.01±14.99g / kg) (P<0.001), and the contents were 249.93±7.45g / kg, 246.39±12.82g / kg and 251.70±6.77g / kg, respectively, while there was no significant difference in the crude protein content between Formula 4 and the control group.

[0046] Crude fat content in fruiting body Figure 3As shown in B, the crude fat content in the fruiting bodies of Formula 3 and Formula 4 (9.37±1.32 g / kg and 8.90±1.31 g / kg) was significantly lower than that in the control group (17.85±0.94 g / kg) (P<0.01), while there was no significant difference in the crude fat content in the fruiting bodies of Formula 1 and Formula 2 compared with the control group.

[0047] according to Figure 3 C shows that only the total sugar content in the fruiting bodies of formula 4 (367.24±39.33 g / kg) was significantly higher than that of the control group (304.06±7.97 g / kg), and there was no significant difference in the total sugar content of the fruiting bodies of the other three formula groups compared with the control group.

[0048] Among the 20 amino acids measured (Table 1), compared with the control group, the contents of 17 amino acids (aspartic acid, glutamic acid, serine, arginine, glycine, threonine, proline, alanine, valine, methionine, cystine, isoleucine, leucine, histidine phenylalanine, lysine and tyrosine) in the fruiting bodies of Formula 1 were significantly higher than those in the control group (P<0.01), while the content of asparagine was significantly lower than that in the control group; the contents of 3 amino acids (aspartic acid, hydroxyproline and serine) in the fruiting bodies of Formula 2 were significantly higher than those in the control group (P<0.01), while the content of alanine was significantly lower than that in the control group; the contents of 2 amino acids (hydroxyproline and proline) in the fruiting bodies of Formula 3 were not significantly different from those in the control group, and the contents of the remaining 18 amino acids were significantly lower than those in the control group (P<0.01); only the content of hydroxyproline in the fruiting bodies of Formula 4 was not significantly different from that in the control group, and the contents of the remaining 19 amino acids were significantly lower than those in the control group (P<0.01). In terms of total amino acid content ( Figure 3 D), Formula 1 was significantly higher than the control group (P<0.001), Formula 3 and Formula 4 were significantly lower than the control group (P<0.001), while Formula 2 had no significant difference with the control group. At the same time, the differences in the content of umami amino acids, bitter amino acids and sweet amino acids in the fruiting bodies of each group were analyzed. Umami amino acids include aspartic acid and glutamic acid. Formula 1 was significantly higher than the control group (P<0.001), and Formula 3 and Formula 4 were significantly lower than the control group (P<0.001) ( Figure 2 E). Sweet amino acids include serine, glycine and threonine, bitter amino acids include arginine, valine and methionine. The contents of sweet amino acids and bitter amino acids in the fruiting bodies of Formula 1 and Formula 2 were significantly higher than those in the control group (P<0.01), while the contents of sweet amino acids and bitter amino acids in the fruiting bodies of Formula 3 and Formula 4 were significantly lower than those in the control group (P<0.001). The above results show that replacing sawdust with a low proportion of longan shells can increase the content of crude protein and multiple amino acids and flavor of shiitake mushroom fruiting bodies, while replacing sawdust with a high proportion of longan shells can increase the total sugar content of fruiting bodies, but reduce the crude fat and amino acid content and flavor of fruiting bodies.

[0049] Table 1 Effects of different longan shell contents on the amino acid content in Lentinus edodes fruiting bodies.

[0050]

[0051]

[0052] Example 3

[0053] Fruiting body samples of the control group, formula one and formula four were collected for volatile metabolome analysis. Take 30±1 mg of the sample in a 2mL EP tube, add 0.24mL of extract solution (methanol-water volume ratio = 3:1), then add 10μL of ribitol, and vortex for 30s; add porcelain beads, treat with a 45Hz grinder for 4min, and ultrasonicate for 5min (ice-water bath); centrifuge the sample at 4℃, and centrifuge at 13000rpm for 15min; carefully transfer 0.18mL of supernatant to a 2mL injection bottle (methyl silyl), take 25μL of each sample and mix them into QC samples; dry the extract in a vacuum concentrator, add 30μL of methoxyamine salt reagent (methoxyamine hydrochloride, dissolved in pyridine 20mg / mL), mix gently, and incubate in an oven at 80℃ for 30min; add 50μL BSTFA (containing 1% TMCS, v / v), incubate the mixture at 70℃ for 1.5h; cool to room temperature, add 5μL of saturated fatty acid methyl ester standard mixture (dissolved in chloroform C8-C16: 1mg / mL; C18-C24: 0.5mg / mL) to the mixed sample; detect in random order on Agilent 7890 gas chromatograph-time of flight mass spectrometer, and use ChromaTOF software (V 4.3x, LECO) to perform peak extraction, baseline correction, deconvolution, peak integration, peak alignment and other analyses on the mass spectrometry data.

[0054] A total of 1019 volatile metabolites were detected in the three groups of Lentinus edodes fruiting body samples. Figure 4A), there were 493 differential metabolites in formula 1 (395 increased and 98 decreased), and there were 521 differential metabolites in formula 4 (421 increased and 100 decreased); compared with formula 1, there were 236 differential metabolites in formula 4 (136 increased and 100 decreased). From 673 differential metabolites, the main volatile aroma substances (eight carbon compounds and sulfur-containing compounds) of 10 kinds of shiitake mushrooms were selected. Among them, 1,2,3,5,6-pentathiacycloheptan (shiitake essence), 1,2,4,6-tetrathiacycloheptan, 1,2,4-trithiacyclopentane, carbonyl sulfide, dimethyl sulfone and methyl mercaptan in the fruiting bodies of formula 1 and formula 4 were higher than those in the control group, while 2,4,5-trithiahexane, methyl propyl disulfide, 3-octanone and 1-octanol were lower than those in the control group; with the increase of the replacement ratio of longan shell, the contents of 2,4,5-trithiahexane, methyl propyl disulfide and 1,2,4,6-tetrathiacycloheptan increased, while the content of 3-octanone decreased ( Figure 4 B). In conclusion, partial replacement of sawdust with longan shells can change the composition of volatile metabolites in Lentinus edodes fruiting bodies and improve the aroma of the fruiting bodies.

[0055] In summary, the cultivation of Lentinus edodes with partial (15% and 30%) replacement of sawdust can promote the growth rate of Lentinus edodes mycelium, increase the content of crude protein, crude fat, total amino acids, umami amino acids, sweet amino acids, bitter amino acids, and improve the quality of Lentinus edodes fruiting bodies; but the high proportion (60%) replacement of sawdust significantly reduces the content of crude protein, crude fat, total amino acids, umami amino acids, sweet amino acids and bitter amino acids in the fruiting bodies. The cultivation medium containing 15% and 60% longan shell increases the content of the main volatile flavor substances in Lentinus edodes fruiting bodies, such as 1,2,3,5,6-pentathiacycloheptan (Mushroom essence), 1,2,4,6-tetrathiacycloheptan, 1,2,4-trithiacyclopentane, carbonyl sulfide and methyl mercaptan.

[0056] Therefore, using longan shells to partially replace sawdust in cultivating shiitake mushrooms can promote mycelium growth and increase the content of nutrients and volatile flavor substances. The present invention provides a preparation method and application of a shiitake mushroom cultivation material formula containing longan shells, which develops a new, relatively green and efficient direction in the field of edible fungus cultivation and agricultural waste recycling.

[0057] The above are only some embodiments of the present invention. However, the present invention is not limited to the above embodiments, and there are many variations. All direct outputs or modified outputs of the content completed under the technical spirit suggested by the present invention should belong to the protection scope of the present invention.

Claims

1. A formula of mushroom cultivation material containing longan shells, characterized in that: Including wood chips, longan shells, bran, gypsum and lime.

2. The mushroom cultivation material formula according to claim 1, characterized in that: Calculated by mass fraction, the longan shell accounts for 15-60%; preferably, the longan shell accounts for 15-45%.

3. The mushroom cultivation material formula according to claim 2, characterized in that: Calculated by mass fraction, the longan shell accounts for 15-30%.

4. The mushroom cultivation material formula according to claim 3, characterized in that: Calculated by mass, it includes 50-65% of sawdust, 15-30% of longan shells, 18.8% of bran, 1% of gypsum and 0.2% of quicklime.

5. The mushroom cultivation material formula according to claim 4, characterized in that: Calculated by mass, it includes 65% sawdust, 15% longan shells, 18.8% bran, 1% gypsum and 0.2% quicklime.

6. A method for preparing a mushroom cultivation material formula containing longan shells according to any one of claims 1 to 5, comprising the following steps: (1) drying longan shells in an oven at 50° C. to 60° C. and crushing them in a crusher; (2) Mix with other planting materials.

7. Application of longan shell in mushroom cultivation.

8. The use according to claim 7, characterized in that: The application of longan shell as a cultivation medium for cultivating shiitake mushrooms can promote the growth of shiitake mushroom mycelium and increase the contents of crude protein, umami amino acid, sweet amino acid, bitter amino acid, total amino acid and shiitake essence in shiitake mushroom fruiting bodies.

9. The use according to claim 8, characterized in that: The invention relates to the use of any one of the mushroom cultivation material formulas in claims 1 to 6 in promoting the growth of mushroom mycelium and increasing the contents of crude protein, crude fat, total amino acids, umami amino acids, sweet amino acids and bitter amino acids in the fruiting bodies, thereby improving the quality of the mushroom fruiting bodies.

10. The use according to claim 7, characterized in that: The invention relates to the application of a mushroom cultivation material formula in increasing the content of main volatile flavor substances in mushroom fruiting bodies. Preferably, the volatile flavor substances are 1,2,3,5,6-pentathiacycloheptane (mushroom essence), 1,2,4,6-tetrathiacycloheptane, 1,2,4-trithiacyclopentane, carbonyl sulfide and methyl mercaptan.

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