A method for efficiently cultivating rhizoma zedoariae and adjusting soil phenolic acid after cultivation

Abstract

The application provides a method for efficiently cultivating rhizoma zyzyphioris and adjusting soil phenolic acid after cultivation. Soil samples are taken at fixed time every month from the time of spore sowing of rhizoma zyzyphioris to the time of fruiting body harvesting. The application analyzes soil physical and chemical properties, soil microbial flora and phenolic acid metabolites. The application optimizes and screens out a more suitable formula for cultivating rhizoma zyzyphioris with miscanthus giganteus and sea mushroom dregs, and discusses the practicability and use form of the secondary use of sea mushroom dregs in the cultivation process of rhizoma zyzyphioris. Finally, the application improves the traditional cultivation formula and mode of rhizoma zyzyphioris in combination with the indexes of soil physical and chemical properties and phenolic acid metabolite content. The technology helps to reduce the adverse effects of rhizoma zyzyphioris cultivation on the increase of soil phenolic acid. The application preliminarily discusses the method of applying the fermented mixture of microbial agent and sea mushroom dregs to the soil for rhizoma zyzyphioris cultivation, which can keep the yield of rhizoma zyzyphioris in the second year at the same level as that in the first year in the same field. The method can provide a reference scheme for relieving the continuous cropping obstacles in the cultivation of rhizoma zyzyphioris.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of agriculture, and particularly relates to a method for efficiently cultivating Dictyophora indusiata and adjusting soil phenolic acid after cultivation. BACKGROUND

[0002] 1. Research significance

[0003] Dictyophora indusiata is a rare edible fungus of the fungal kingdom, the Dictyophoraceae family and the Dictyophora genus, which contains various enzymes and high-molecular polysaccharides. With the increasing demand for Dictyophora indusiata, the cultivation industry of Dictyophora indusiata has been rapidly developed in Fujian, Yunnan and Guizhou provinces, and the cultivation scale and yield of Dictyophora indusiata have been continuously improved. Cultivation of Dictyophora indusiata has become an important economic source for local farmers.

[0004] Traditional cultivation formula of Dictyophora indusiata requires a large amount of bamboo chips and sawdust, which has a high production cost, a limited source and a great impact on the ecological environment. Therefore, carrying out edible fungus cultivation by using grass instead of wood is an important technical means to alleviate the contradiction between fungus and forest. Shunchang County is a major production area of Dictyophora indusiata, and tons of waste Dictyophora indusiata dregs are produced every day. Studies have found that the waste Dictyophora indusiata dregs contain a large amount of fiber, lignin and many trace elements, and can still be used for substitute material cultivation of other edible fungi. Reasonable use of waste Dictyophora indusiata dregs can not only save production cost, but also effectively reduce environmental pollution caused by improper disposal of dregs. At present, Mao Ling et al. have successfully cultivated edible fungi such as Phellinus baumii, Agrocybe aegerita and Volvariella volvacea by using waste Dictyophora indusiata dregs as substitute material.

[0005] For the edible fungus cultivated by covering soil, it is found that the activity of Agaricus bisporus mycelium gradually decreases with the decrease of laccase activity, while the activity of Ascomycota laccase increases after the first flush of mushrooms. Studies on possible allelopathic autotoxic substances show that these chemical substances generally affect the soil enzyme activity and nutrient metabolism of crops to inhibit the growth and development of crops.

[0006] Therefore, in the present application, giant reed and waste Dictyophora indusiata dregs are used to partially replace traditional cultivation material. The effects of giant reed and waste Dictyophora indusiata dregs on cultivation of Dictyophora indusiata and soil microecology after cultivation are analyzed from the aspects of soil physical and chemical properties, soil microbial flora and phenolic acid metabolites, so as to comprehensively analyze the soil condition after cultivation of Dictyophora indusiata by taking soil microorganisms and phenolic acid metabolites as key points.

[0007] 2. Research status at home and abroad

[0008] 2.1 Edible fungus cultivation technology by using grass

[0009] Cenchrus fungigraminus is rich in crude protein and crude fiber, which are the nutrients needed for edible mushroom cultivation materials. It has the characteristics of high yield per mu and the advantage of developing "grass instead of wood". In the long-term exploration and practice, it has successfully cultivated many edible and medicinal mushrooms including shiitake, agaric, bamboo, and monkey head mushroom. Through mushroom cultivation, not only can a large amount of edible mushrooms be produced efficiently in a short time, but also the nutritional value of the cultivated mushrooms can be improved. At the same time, the cost of mushroom cultivation is relatively low, which can effectively reduce the production cost.

[0010] In the formula screening of Cenchrus fungigraminus cultivation of edible mushrooms, it was found that the best ratio of Cenchrus fungigraminus and cottonseed hulls was 1:1 in the formula screening experiment of Pleurotus ostreatus. The biological conversion efficiency and input-output ratio of different formulas showed that Cenchrus fungigraminus could be used as the main raw material for cultivating Pleurotus ostreatus. Dong Xiaona et al. found that the mycelial growth rate was proportional to the content of Cenchrus fungigraminus when screening the best formula for Cenchrus fungigraminus cultivation of Ganoderma lucidum. However, considering the mycelial growth rate, mycelial growth potential, and yield of the mushroom bag, the optimal formula was 48% Cenchrus fungigraminus.

[0011] In summary, Cenchrus fungigraminus cultivation of edible mushrooms has the advantages of protecting the ecological environment, reducing costs, realizing resource recycling, and high efficiency and yield.

[0012] 2.2 Secondary utilization of edible mushroom residues

[0013] Mushroom residues, also known as mushroom dregs, are the cultivation materials used for growing edible mushrooms. During the growth of mycelium, extracellular enzymes are secreted to degrade lignin, cellulose, and other substances in the cultivation material. After the fruiting bodies are harvested, the remaining residues are left behind. China is the world's largest producer, consumer, and exporter of edible mushrooms. According to reports, it is estimated that 5 kg of mushroom residues are produced for every 1 kg of edible mushrooms. If not properly utilized, mushroom residues can be randomly discarded or burned by farmers, resulting in resource waste and air pollution. However, through relevant research and promotion, mushroom residues are no longer considered waste, but renewable resources that have been widely used in agriculture. The main ways of resource utilization include: making edible mushroom cultivation media, producing organic fertilizers, making fertilizers, and adsorbing soil pollutants.

[0014] The physicochemical properties and nutritional components of different spent mushroom composts are different due to different cultivated species and raw materials. The spent mushroom compost of Pleurotus cornucopiae has the characteristics of high contents of soluble protein, reducing sugar and starch, and the content of crude polysaccharide is as high as 33.40 mg / g. The research on the effects of the extract of the spent mushroom compost of Pleurotus cornucopiae on the growth of the mycelium of Tricholoma giganteum and the activity of laccase shows that the extract increases the growth speed, growth amount and laccase activity of the mycelium of Tricholoma giganteum, and is a suitable cultivation raw material for Tricholoma giganteum. In addition, in the research on the effects of the spent mushroom compost of Agaricus bisporus on the seedling effect of tomatoes, it is found that no matter what the concentration of the spent mushroom compost is, the addition of the spent mushroom compost can improve the quality of the tomato seedlings compared with the vermiculite group, and when the ratio of the spent mushroom compost to the vermiculite is 1:1, the quality of the tomato seedlings and the quality of the cultivation substrate are the best. Therefore, the spent mushroom compost can be used as an agricultural production resource to provide nutrients for crops and maximize the utilization of resources.

[0015] 2.3 Effects of cultivation of edible fungi on soil phenolic acids

[0016] Phenolic acids are one of the most abundant secondary metabolites in higher plants and an important class of plant allelochemicals. They can enter the soil environment through root exudation, decomposition of residues and litter, and rain and mist leaching. Studies have shown that when phenolic acids accumulate to a certain concentration in the soil, they can inhibit root growth, reduce root enzyme activity, affect hormone synthesis and photosynthesis, and even cause poor growth, disease and death. Some studies have also found that phenolic acids can promote the proliferation of soil-borne pathogens, disrupt the balance of soil microecology, and increase the risk of crop soil-borne diseases and continuous cropping obstacles. Therefore, phenolic acids are considered to be the key inducers driving the occurrence of continuous cropping obstacles in crops. In the study of the cultivation of Rhizoma Zanthorosae, Wu Hongping et al. found that the continuous cropping of Rhizoma Zanthorosae under rubber plantations affected the soil physical and chemical properties and soil enzyme activity, and the accumulation of "toxic by-products" produced by the "autolysis" of Rhizoma Zanthorosae affected the number and function of cultivable microorganisms and functional microorganisms in the soil, and also affected the functional characteristics of soil microbial communities. Therefore, changes in the "microenvironment" of the soil may be the main reason for the continuous cropping obstacles of Rhizoma Zanthorosae under rubber plantations.

[0017] The present application can provide scientific basis for the cultivation of Rhizoma Zanthorosae, optimize the production and management measures of Rhizoma Zanthorosae, improve the yield and quality of Rhizoma Zanthorosae, reduce the occurrence of diseases and pests, and protect the ecological environment of the soil by optimizing the soil microecology and culture medium of Rhizoma Zanthorosae cultivation, and applying the spent mushroom compost. SUMMARY

[0018] The present application aims to provide a method of using giant reed and spent mushroom compost of Pleurotus cornucopiae for joint fermentation and application, to improve the yield and quality of Rhizoma Zanthorosae, and to reduce the content of phenolic acids in the soil after the cultivation of Rhizoma Zanthorosae.

[0019] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0020] A method for efficiently cultivating bamboo shoots and reducing soil phenolic acid after cultivation, comprising the following steps:

[0021] 1. Cultivation pad: collect fresh, disease-free sea mushroom dregs (90wt%), after crushing, mixed with urea (5wt%), light calcium carbonate (5wt%), and then stacked and fermented according to the size of 1.4-1.6 meters high, 1.4-1.6 meters wide, and unlimited length, and uniformly turned over once about 10 days, so that the sea mushroom dregs are uniformly fermented, the moisture content is 60-65tw%, and the fermentation time is 35-45 days;

[0022] 2. Cultivation formula: mix giant grass and sawdust according to the mass ratio of 1:1, and then stack and ferment according to the size of 1.4-1.6 meters high, 1.4-1.6 meters wide, and unlimited length, and uniformly turn over once about 10 days, so that the sawdust and giant grass are uniformly mixed and fermented, the moisture content is 60-70wt%, and the fermentation time is 40-45 days;

[0023] 3. Application method: during the cultivation of bamboo shoots, the fermented sea mushroom dregs are used as the pad with a height of about 10cm (the pad usage is 2.5 tons per mu), the fermented giant grass and sawdust are used as the main culture medium, and the bamboo shoots are divided into the size of an egg and inoculated at a distance of 30-50cm each, and then covered with soil with a height of 3-5cm, the usage of bamboo shoot spores is 200-230 bags per mu, about 0.9kg per bag, and the bamboo shoot mycelium recovery and fruiting body cultivation stages are carried out, and the management method and mode of the bamboo shoot fruiting body are consistent with the traditional way (controlling the relative humidity of air to be 85%-95%, and only scattering light is needed during the growth stage of the fruiting body);

[0024] 4. It is determined that the yield and quality of the bamboo shoot fruiting body cultivated by the method are higher than those of the traditional cultivation method.

[0025] 5. Through high performance liquid chromatography mass spectrometry analysis, the content of total soil phenolic acid (ferulic acid, coumaric acid, p-coumaric acid, syringic acid, vanillic acid and p-hydroxybenzoic acid) during the cultivation of the bamboo shoot by the method is significantly lower than that of other formulas and cultivation modes, which helps to reduce the impact of bamboo shoot cultivation on the soil.

[0026] 6. The soil conditioner of the combination of bacillus subtilis agent (1kg per mu) and fermented sea mushroom dregs (6 tons per mu) is applied to the soil once in the third month before the cultivation of bamboo shoots, and the yield per mu basically recovers to be consistent with the first year in the second year of continuous cultivation.

[0027] Compared with the prior art, the present application has the following advantages:

[0028] 1. At present, there is a fungus as soil fertilizer to increase soil organic matter, total nitrogen and other effects, but there is no related report on whether it affects the content of phenolic acid in the soil after planting bamboo.

[0029] 2. The traditional cultivation of bamboo mainly uses sawdust and bamboo chips, and the present application uses giant grass and sea mushroom fungus dregs as the main raw material to cultivate bamboo, and can obtain stable and high yield of bamboo fruiting bodies.

[0030] 3. The formation time of bamboo fungus egg is obviously shortened (5-7 days), and the mushroom time of bamboo fruiting body is prolonged (20-35 days). BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 The first year of the experimental treatment diagram;

[0032] Figure 2 The content of phenolic acid in the soil of different treatment groups at each stage; note: a is the accumulation column chart of the content of phenolic acid in the soil of different treatment groups at each stage; b is the percentage column chart of the content of phenolic acid in the soil of different treatment groups at each stage. R-JS in the figure is fungus dregs-substrate of fungus grass sawdust group, R-JR is fungus dregs-substrate of fungus grass fungus dregs group, JS is fungus grass sawdust group, JR is fungus grass fungus dregs group. CK represents the soil sample before planting, Apr, May and Jun represent the soil samples in April, May and June respectively.

[0033] Figure 3 PCoA analysis of soil microbial diversity of different experimental groups, (a) is the PCoA analysis of fungal community; (b) is the PCoA analysis of bacterial community; note: R-JS in the figure is 50% giant grass and 50% sawdust as cultivation material, sea mushroom fermented fungus dregs as bedding experimental group, R-JR is 50% giant grass and 50% sea mushroom fungus dregs as cultivation material, sea mushroom fermented fungus dregs as bedding experimental group, JS is 50% giant grass and 50% sawdust as cultivation material experimental group, JR is 50% giant grass and 50% sea mushroom fungus dregs as cultivation material experimental group; CK represents the soil sample before planting. DETAILED DESCRIPTION

[0034] A method for efficiently cultivating bamboo and reducing the content of phenolic acid in soil after cultivation, comprising the following steps:

[0035] 1. Cultivation bedding: collect fresh, disease-free sea mushroom fungus dregs (90wt%), crush and mix with urea (5wt%) and light calcium carbonate (5wt%), then stack and ferment according to the size of 1.5 meters high, 1.5 meters wide and 3 meters long, and uniformly turn over once about 10 days to make the sea mushroom fungus dregs uniformly fermented, with a moisture content of 60wt%, and a fermentation time of 40 days;

[0036] 2. Cultivation formula: Ferment the giant grass and sawdust according to a mass ratio of 1:1 by stacking them in a cycle of one layer of giant grass and one layer of sawdust to form a pile 1.5 meters high, 1.5 meters wide, and 3 meters long. Ferment for about 10 days, and uniformly turn over the pile once to mix the sawdust and giant grass evenly. The water content is 65 wt%, and the fermentation time is 40 days.

[0037] 3. Application method: During the cultivation of bamboo lotus, ferment the mushroom dregs as a bedding material about 10 cm high (the bedding material dosage is 2.5 tons per mu), and use the fermented mushroom grass and sawdust as the main culture medium. Pile the material according to a specification of 40 cm high and 60 cm wide. Divide the bamboo lotus into egg-sized pieces and inoculate them at a distance of 30 cm each, then cover them with soil 3 cm high. The bamboo lotus spore dosage is 200 bags per mu, about 0.9 kg per bag. Perform the bamboo lotus mycelium recovery and fruiting body cultivation stages. The bamboo lotus fruiting body management method and mode are consistent with the traditional method (control the air relative humidity to 90%, and maintain the light intensity at 1000 Lux during the growth of the fruiting body).

[0038] 4. Use the Bacillus subtilis agent (1 kg per mu) and the fermented mushroom dregs (6 tons per mu) to jointly ferment the soil modifier. Apply it to the soil once in the third month before cultivating bamboo lotus. The yield per mu basically recovers to be consistent with the first year in the second year of continuous cultivation.

[0039] 1. Method and steps

[0040] 1.1 Test design

[0041] The field bamboo lotus cultivation experiment was carried out from December 2022 to July 2024 in the Giant Grass Technology Courtyard Test Area in Xilan Village, Shunchang County, Nanping City, Fujian Province (26°45'7"N, 117°48'20"E). It is located in the subtropical monsoon climate zone with mild and humid climate, abundant rainfall, distinct seasons, short winter, long summer, early spring, and late autumn. The average annual temperature is 18.5°C, the average annual rainfall is 1756 mm, and the average annual sunshine is 1740.7 hours. In the first year of the experiment, according to the different culture medium formulas, it was divided into four groups, namely mushroom dregs-giant grass + sawdust (R-JS), mushroom dregs-giant grass + mushroom dregs (R-JR), giant grass + sawdust (JS), and giant grass + mushroom dregs (JR). Each group was repeated three times in the field. The specific layout is as follows: Figure 1The soil amendment was prepared by Bacillus subtilis combined with fermented Panus conchatus vinasse in the second year experiment. The soil of the first year of Rhizoma zyrianthae cultivation (vinasse substrate-fermented Panus conchatus vinasse + sawdust (R-JS) group) was applied with the soil amendment three months in advance (R-3), two months in advance (R-2), one month in advance (R-1) and without application (CK). The six-point sampling method was used to sample the soil at 0-15 cm under the Rhizoma zyrianthae cultivation substrate in the early month of each month during the two years, from the Rhizoma zyrianthae cultivation before the mycelium seeding to the generation of the first flush of fruiting bodies, using the cutting ring method, and then collected in sterile bags, stored in dry ice and brought back to the laboratory. After the samples were collected, the samples for soil physical and chemical properties, enzyme activity and phenolic acid compound analysis were spread in a cool and ventilated place, dried after air drying, sieved and sealed for storage before testing; the samples for microbial diversity analysis were stored in a-80°C refrigerator for testing.

[0042] The above-mentioned straw was selected as giant reed, the vinasse substrate was fermented Panus conchatus vinasse, and the sawdust was selected as general market mixed sawdust.

[0043] 1.2 Analysis of Rhizoma zyrianthae yield and fruiting body agronomic traits

[0044] The total yield of Rhizoma zyrianthae fruiting bodies in all treatments was collected; the diameters of Rhizoma zyrianthae sclerotia, fruiting body stem length, stem diameter, cap length and other agronomic traits of Rhizoma zyrianthae sclerotia in different treatment groups were analyzed from the formation of Rhizoma zyrianthae sclerotia. The picked fruiting bodies were stored at low temperature and transported back to the laboratory, the samples were dried to constant weight using an oven to determine the moisture content, and GB / T15672-2009 was used for total sugar determination.

[0045] 1.3 Analysis of soil physical and chemical properties

[0046] The Kjeldahl method was used to determine the total nitrogen (TN) of the soil; the alkali fusion-molybdenum antimony anti-spectrophotometric method was used to determine the total phosphorus (TP) of the soil; the NAOH fusion-flame photometry method was used to determine the total potassium (TK); the potassium dichromate oxidation-spectrophotometric method was used to determine the soil organic carbon (OC); the soil organic matter (OM) was calculated through the conversion coefficient of organic carbon and organic matter; the KCl extraction-indigo phenol blue colorimetric method was used to determine the soil ammonium nitrogen (AMN); the nitro salicylic acid method was used to determine the soil nitrate nitrogen (NIN); the potential method was used to determine the soil pH.

[0047] 1.4 Analysis of soil enzyme activity

[0048] The 3,5-dinitrosalicylic acid colorimetric method was used to determine the soil sucrose (S-SC); the nitrophenyl phosphate disodium colorimetric method was used to determine the soil acid phosphatase (S-ACP); the 2,2-diazene-bis (3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt (ABTS) colorimetric method was used to determine the soil laccase (S-L); the potassium permanganate titration method was used to determine the soil catalase (S-CAT); the indigo phenol colorimetric method was used to determine the soil urease (S-UE).

[0049] 1.5 Soil phenolic acid content determination

[0050] Take 10 g of soil sample into a 50 mL centrifuge tube, add 25 mL of 2 mol / L NAOH solution, stand for 24 h, then place in a 200 r / min, 25°C shaking bed and shake for 2 h. Centrifuge at room temperature at 8000 r / min for 15 min, take 15 mL of supernatant, adjust pH to 2.5 using 36% HC1 solution to remove humic acid. Centrifuge at 8000 r / min for 15 min, take 15 mL of supernatant, extract with equal amount of ethyl acetate three times, combine the extract. The extract is rotary evaporated to dryness at 45°C, redissolved with 2 mL of methanol and filtered through a 0.22 μm filter for testing. Use a Waters high performance liquid chromatography system (E2695, Waters, Milford, USA) with a diode array detector, use Uranus C18 column (250 mm x 4.6 mm x 5 μm) and guard column (20 mm x 4.6 mm x 5 μm) for detection analysis at 280 nm. Use the following mobile phase solution for high performance liquid chromatography separation: trifluoroacetic acid aqueous solution (volume fraction 0.1%) and acetonitrile mobile phase, column temperature 30°C, injection volume 10 μL, flow rate 1.0 mL / min, elution gradient is 5%-10% acetonitrile (0-10 min), 10%-10% acetonitrile (10-20 min), 10%-35% acetonitrile (20-60 min), 35%-60% acetonitrile (60-75 min), 60%-60% acetonitrile (75-85 min), 60%-5% acetonitrile (85-86 min), 5%-5% acetonitrile (86-90 min) and keep for 5 min. Each injection interval is column equilibration with mobile phase for 10 min.

[0051] 1.6 Diversity analysis of soil microorganisms in bamboo shoot cultivation

[0052] DNA of soil microorganism in each experimental group in field experiment was extracted using the kit of American MP Biomedicals Company (MagBeads FastDNA Kit for Soil), and after the concentration and quality of DNA reached the standard, PCR (2720, ABI, USA) amplification was carried out. The diluted genomic DNA was used as a template, and the barcode-carrying primer was used for amplification and quantification. The bacterial genome (16S rDNA V3-V4 region) primer was 338F (5'-ACTCCTACGGGAGGCAGCA-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3'). The fungal genome (ITS1) primer was ITS1F (5'-GGAAGTAAAAGTCGTAACAAGG-3') and ITS2R (5'-GCTGCGTTCTTCATCGATGC-3'). After the purified product was connected to the sequencing adapter, the sequencing library was constructed, and the community DNA fragments were double-end sequenced using the Illumina platform.

[0053] After splicing, filtering and removing chimeras, the primer fragments of the sequences were removed using qiime2 software (Version 2019.4) in the call qiimecutadapt trim-paired, and the sequences that did not match the primer were discarded; then DADA2 was called by qiimedada2 denoise-paired for quality control, denoising, splicing and chimeric removal. The Qiime2 software (Version 2019.4) was used for species annotation by referring to the Unite database using the classify-sklearn algorithm. In the Alpha diversity analysis, Chao1 and Observed species index were used to represent richness, Shannon and Simpson index were used to represent diversity, Faith's PD index was used to represent evolution-based diversity, Pielou's evenness index was used to represent uniformity, and Good's coverage index was used to represent coverage. In the Beta diversity analysis, principal coordinate analysis (PCoA) was used for dimensionality reduction and sorting analysis.

[0054] 2. Results and analysis

[0055] 2.1 Effects of different treatments on the yield and quality of Dictyophora indusiata fruiting bodies

[0056] From Table 1, the water content of bamboo shoot fruit bodies of different treatment groups has no significant difference (P<0.05). The single weight of bamboo shoot fruit bodies arranged from large to small is R-JR>JS>JR>R-JS, which is 24.57 g, 21.33 g, 21.15 g and 12.88 g respectively. The fresh weight of bamboo shoot fruit bodies per mu arranged from large to small is R-JR>JS>R-JR>JR, which is 1406.25 kg·mu -1 , 1271.25 kg·mu -1 , 896.25 kg·mu -1 , 708.75 kg·mu -1 . The diameter of the spore and the diameter of the stipe of different treatment groups have no significant difference. Whether there is padding in the JS group has significant difference in the length of the cap and the length of the stipe, and there is no significant difference in the JR group. In the two formulations using distiller's grains padding, the length of the cap has no significant difference, and the length of the stipe has no significant difference. In the two formulations without distiller's grains padding, the length of the cap has significant difference, and the length of the stipe has no significant difference.

[0057] From Table 2, the total sugar content of bamboo shoots arranged from large to small is JS>R-JS>JR>R-JR, which is 61.72%, 58.31%, 50.93% and 37.19% respectively. Among them, there is no significant difference between JS group with and without distiller's grains, and there is significant difference in JR group. There is significant difference between different treatment groups with and without distiller's grains padding, that is, there is significant difference between R-JS and R-JR, and there is significant difference between JS and JR groups.

[0058] Table 1 Bamboo shoot agronomic traits

[0059]

[0060] Note: In the table, R-JS is distiller's grains bedding-grass sawdust group, R-JR is distiller's grains bedding-grass distiller's grains group, JS is grass sawdust group, and JR is grass distiller's grains group. The content of each index is represented by mean ± standard deviation. According to ANOVA single factor variance analysis, the same column data after different lowercase letters represent the degree of difference (p<0.05).

[0061] Table 2 Nutritional composition of bamboo shoots

[0062]

[0063]

[0064] Note: R-JS is the fungus dregs-wood chip group, R-JR is the fungus dregs-fungus dregs group, JS is the fungus wood chip group, and JR is the fungus dregs group. The index content is expressed as the mean ± standard deviation. According to ANOVA single factor variance analysis, different lowercase letters in the same column indicate the degree of significance (p < 0.05).

[0065] 2.2 Analysis of the physical and chemical properties of the soil for the cultivation of Rhizoma Zephyranthes

[0066] The changes in the physical and chemical properties of the soil during the growth period of Rhizoma Zephyranthes in each experimental group are shown in Table 3. In the R-JS group, the six indicators except pH and AMN showed a trend of first increasing, then decreasing, and then increasing again. The TN, NIN, OC, and OM of the September samples increased by 36.81%, 22.50%, 12.05%, 62.97%, 44.53%, and 44.53%, respectively, compared to CK, and the pH and AMN decreased by 5.68% and 23.22%, respectively. In the R-JR group, the TN, OC, and OM indicators showed a trend of first increasing, then decreasing, and then increasing again. The above four indicators increased by 26.82%, 10.29%, 36.63%, and 36.62%, respectively, in September compared to CK. The AMN showed a trend of first increasing and then decreasing, and decreased by 18.52% and 9.24% in September compared to CK. The NIN showed a trend of first decreasing and then increasing, and significantly decreased from 2.288 μg / mL to 0.684 μg / mL in May compared to CK. The pH decreased from 6.07 to 6.01. In the JR group, the TN, AMN, NIN, OC, OM, and pH indicators showed a trend of first increasing, then decreasing, and then increasing again. The above six indicators increased by 4.56%, 28.30%, 48.79%, 20.84%, 20.84%, and 14.58%, respectively, in September compared to CK, and the NIN significantly increased to 5.443 μg / mL in May. The pH increased significantly from 6.07 to 7.11. In the JS group, the TN, NIN, OC, OM, and pH indicators showed a trend of first increasing, then decreasing, and then increasing again. The above five indicators increased by 17.87%, 48.79%, 29.73%, 29.73%, and 1.93%, respectively, in September compared to CK.

[0067] Only compared CK and the end of the mushroom picking period, i.e. September, two bamboo shoots growth period found that, except JS group, soil total nitrogen was significantly increased, and in the end of mushroom picking period, i.e. September, the soil total nitrogen content of bedding group was higher than that of no bedding group, and the content from large to small was R-JS > R-JR > JR > JS; soil ammonium nitrogen was reduced except JS group, and the content from large to small was JS > R-JR > R-JS > JR; soil nitrate nitrogen was increased, and the content from large to small was R-JS > JS > JR > R-JR; the soil pH of bedding group, i.e. R-JS and R-JR, was reduced, and the soil pH of no bedding group, i.e. JS and JR, was significantly increased, and in the same formula, the soil pH of no bedding group was higher than that of bedding group, and the content from large to small was JS > JR > R-JR > R-JS; soil organic matter and organic carbon content were significantly increased, and in the same formula, the soil organic matter and organic carbon content of bedding group was higher than that of no bedding group.

[0068] Table 3 Soil physicochemical properties of different experimental groups at different periods

[0069]

[0070] Note: R-JS in the table is straw bedding-straw sawdust group, R-JR is straw bedding-straw bedding group, JS is straw sawdust group, JR is straw bedding group; CK represents soil sample before planting, Apr, May, Jun, Sept are April, May, June, September soil samples. The content of each index is represented by mean ± standard deviation, according to ANOVA single factor variance analysis, the same column data after different lowercase letters represent the degree of difference (p < 0.05).

[0071] 2.3 Soil enzyme activity of bamboo shoots

[0072] The change trend of soil physicochemical properties in each experimental group during the growth period of Dictyophora was shown in Table 4. In the R-JS group, the activities of S-ACP, S-SC and S-L first increased, then decreased, and then increased again, and the activities of these three enzymes in the soil in September, the last mushroom picking period, were significantly higher than those in the CK soil before planting, increasing by 29.25%, 90.12% and 10.03%, respectively. The activities of S-CAT and S-UE first increased and then decreased, and the activities of these two enzymes in the soil in September were lower than those in the CK, decreasing by 65.28% and 25.71%, respectively. Compared with the soil samples in June, the first mushroom picking period, and the soil samples in September, the last mushroom picking period, it was found that the activities of S-ACP and S-L in the soil in September were significantly higher than those in June, and the activities of S-CAT and S-UE in the soil in September were significantly lower than those in June. The results showed that planting Dictyophora in the R-JS group promoted the activities of S-CAP and S-L in the soil, and inhibited the activities of S-CAT and S-UE. In the R-JR group, the activity of S-ACP in the soil in September, the last mushroom picking period, was significantly higher than that in the CK soil before planting, increasing by 30.73%, and the activities of S-CAT and S-L were significantly lower than those in the CK, decreasing by 39.57% and 13.48%, respectively. Compared with the soil samples in June, the first mushroom picking period, and the soil samples in September, the last mushroom picking period, it was found that the activity of S-ACP in the soil in September was significantly higher than that in June, and the activities of S-CAT and S-L in the soil in September were significantly lower than those in June. The results of planting showed that planting Dictyophora in the R-JR group promoted the activities of S-ACP and S-SC in the soil, and inhibited the activities of S-CAT, S-L and S-UE. In the JS group, the activity of S-ACP in the soil in September, the last mushroom picking period, was higher than that in the CK soil before planting, increasing by 33.01%, and the activities of S-CAT, S-SC, S-L and S-UE were lower than those in the CK, decreasing by 15.70%, 51.48%, 41.53% and 28.24%, respectively. Compared with the soil samples in June, the first mushroom picking period, and the soil samples in September, the last mushroom picking period, it was found that the activity of S-ACP in the soil in September was significantly higher than that in June, and the activity of S-CAT in the soil in September was significantly lower than that in June. The results of planting showed that planting Dictyophora in the JS group promoted the activity of S-ACP in the soil, and inhibited the activities of S-CAT, S-SC, S-L and S-UE. In the JR group, the activity of S-ACP in the soil in September, the last mushroom picking period, was significantly higher than that in the CK soil before planting, increasing by 32.36%, and the activity of S-UE was significantly lower than that in the CK, decreasing by 20.90%. Compared with the soil samples in June, the first mushroom picking period, and the soil samples in September, the last mushroom picking period, it was found that the activities of S-CAT and S-UE in the soil in September were significantly lower than those in June. The results of planting showed that planting Dictyophora in the JR group promoted the activity of S-ACP in the soil, and inhibited the activities of S-CAT, S-SC, S-L and S-UE.

[0073] Compared with the other groups, the S-ACP activity of the bedding groups, i.e. R-JS and R-JR groups, showed a trend of first increasing, then decreasing, and then increasing again, while the S-ACP activity of the non-bedding groups, i.e. JS and JR groups, showed a trend of first decreasing, then increasing. The bedding treatment had a greater impact on S-ACP activity in the first month after mycelium inoculation, i.e. April, and the S-ACP activity of the bedding groups was higher than that of the non-bedding groups. In S-CAT, the activity of the nine-month soil samples of the three groups except JR group was significantly lower than that of CK, and the activity of the non-bedding groups was higher than that of the bedding groups. The activity of JR formula was higher than that of JS group. The results showed that appropriate use of spent grains could effectively promote S-CAT activity, and excessive use would inhibit S-CAT activity. Except for the JS group, the S-SC activity in September was higher than that of CK, and the order from large to small was R-JS > JR > R-JR > JS. Spent grains showed a low-promotion-high-inhibition effect on S-SC activity. The S-L activity showed a downward trend except for the R-JS group, and the S-L activity of the bedding groups was higher than that of the non-bedding groups in April. Compared with the non-bedding groups, the S-L activity of JR group was higher than that of JS group in the first and last mushroom picking periods. The S-UE in September was lower than that of CK. Compared with R-JS and JS, adding spent grain bedding could increase S-UE activity in the first mushroom picking period, i.e. June. Compared with JS and JR, increasing spent grain in the cultivation material could increase S-UE activity in the first mushroom picking period, i.e. June.

[0074] Table 4 Soil enzyme activities of different experimental groups at different periods

[0075]

[0076]

[0077] Note: R-JS in the table is spent grain substrate-straw sawdust group, R-JR is spent grain substrate-straw spent grain group, JS is straw sawdust group, and JR is straw spent grain group. CK represents the soil sample before planting, Apr, May, Jun, and Sept represent April, May, June, and September soil samples. The average value ± standard deviation is used to represent the content of each index. According to ANOVA single factor variance analysis, different lowercase letters after the same column data represent the degree of difference (p < 0.05).

[0078] 2.4 Analysis of the diversity of soil microorganisms in Dioscorea batatas cultivation soil treated with different soil improvers

[0079] The effects of different experimental groups on the fungal diversity in the soil for cultivating Rhizoma Dioscoreae were shown in Table 5. The Chao1, Observed-species, Shannon and Simpson indexes in the soil before cultivating Rhizoma Dioscoreae were higher than those in the four experimental groups, and there were significant differences between R-JS, R-JR and JS. However, there was no significant difference between Chao1 and Observed-species indexes in JR group, and there was a significant difference between Shannon and Simpson indexes in JR group. Among the four experimental groups, the Chao1, Observed-species, Shannon and Simpson indexes in JR group were significantly higher than those in R-JS, R-JR and JS groups. The Chao1 and Observed-species indexes in JS group were the lowest and significantly lower than those in the two experimental groups using distiller's grains as the bedding. The results showed that different cultivation formulas could reduce the community species richness, diversity and evenness in the soil for cultivating Rhizoma Dioscoreae. The analysis of the four indexes showed that the four experimental groups were ranked in descending order as JR > R-JR > R-JS > JS in Chao1 and Observed-species indexes, JR > R-JS > JS > R-JR in Shannon index, and JR > R-JS > JS > R-JR in Simpson index. The results showed that using distiller's grains in the cultivation material or bedding could alleviate the decrease in fungal diversity in the soil after cultivating Rhizoma Dioscoreae.

[0080] The effects of different experimental groups on the bacterial community diversity in the soil for cultivating Rhizoma Dioscoreae were shown in Table 5. The Chao1 and Observed-species indexes in the four experimental groups were ranked in descending order as R-JR > CK > R-JS > JS > JR, and the Shannon index was ranked in descending order as R-JR > JS > CK > JR > R-JS. The results showed that when using "jumbo grass + sawdust" as the cultivation material, the treatment without increasing distiller's grains bedding could better increase the bacterial community species richness and diversity; when using "jumbo grass + distiller's grains" as the cultivation material, the treatment with increasing distiller's grains bedding could better increase the bacterial community species richness and diversity. Moreover, the indexes in R-JR group were always greater than those in CK, indicating that adding distiller's grains in the cultivation material and using distiller's grains bedding could alleviate the decrease in bacterial community species diversity in the soil.

[0081] Principal coordinate analysis (PCoA) was performed to analyze the diversity and heterogeneity of microbial communities using the Bray-Curtis distance selection algorithm. Based on the principle that closer projected distances on the coordinate axes indicate greater similarity in community composition across corresponding dimensions, and using the Permanova test, analysis of the CK and the four experimental groups revealed highly significant differences in fungal community composition between the CK and the four experimental groups, as well as among the four experimental groups themselves: PCoA1 was 35.7%, and PCoA2 was 14.2%. Figure 3 (a) Significant differences were found in the bacterial community composition between the CK and the four experimental groups, as well as among the four experimental groups: PCoA1 was 20.8%, and PCoA2 was 12.9%. Figure 3 (b) The results showed that the soil microbial community diversity changed significantly after cultivating bamboo fungus.

[0082] Table 5. Effects of different experimental groups on soil fungal diversity

[0083]

[0084] Note: In the table, CK represents the soil sample before planting; R-JS represents the mushroom substrate-mushroom grass and sawdust group; R-JR represents the mushroom substrate-mushroom grass and mushroom substrate group; JS represents the mushroom grass and sawdust group; and JR represents the mushroom grass and mushroom substrate group. The content of each indicator is expressed as mean ± standard deviation. According to ANOVA one-way ANOVA, different lowercase letters after the data in the same column indicate the significance of the difference (p<0.05).

[0085] 2.5 Effects of different treatments on soil phenolic acid

[0086] Based on the results of high-performance liquid chromatography (HPLC), a standard curve was plotted with the concentrations of six phenolic acid standards on the x-axis and the peak area integral on the y-axis. The obtained linear equation and regression coefficient R0 were then analyzed. 2 As shown in Table 6.

[0087] Table 6 Linear equations for phenolic acid standards

[0088]

[0089]

[0090] Based on the results of high-performance liquid chromatography (HPLC), the measured peak areas were substituted into the linear equation of the aforementioned standards to calculate the concentration of phenolic acids in different soil samples. The changes in total phenolic acid content in different groups before bamboo fungus cultivation, after mycelial inoculation, and during the first ten days of each month after the first flush of fruiting bodies are shown below. Figure 2 As shown, the total phenolic acid content in the soil before bamboo fungus cultivation was 3.807 mg·kg⁻¹. -1, the total phenolic acid content of the first fruiting body of the four treatment groups showed a trend of first increasing and then decreasing, and the total phenolic acid content of the first fruiting body of the R-JS group, the R-JR group, the JS group and the JR group was 3.517 mg·kg -1 , 4.715 mg·kg -1 , 5.017 mg·kg -1 , 5.998 mg·kg -1 , respectively. The content of the JR group was the highest, and the content of the R-JS group was the lowest. Compared with CK, the total phenolic acid content of the first fruiting body of the R-JS group decreased by 8.22%, and the total phenolic acid content of the first fruiting body of the R-JR, JS and JR groups increased by 19.27%, 24.12% and 36.54%, respectively.

[0091] 3. Conclusion

[0092] Using fermented seafood mushroom dregs as bedding, cultivating bamboo shoots with giant bacteria grass and sawdust can significantly improve the yield and variety of bamboo shoot fruiting bodies, stabilize the phenolic acid content in bamboo shoot cultivation soil, reduce the harm of bamboo shoot cultivation to the soil; at the same time, in the second year, using Bacillus subtilis and seafood mushroom dregs fermented soil conditioner to treat the soil three months before cultivation can effectively alleviate the continuous cropping obstacles of bamboo shoots and provide scientific reference.

[0093] The above is only the preferred embodiment of the present application, and any equivalent changes and modifications made within the scope of the present application are also included in the scope of the present application.

Claims

1. A method for efficient cultivation of bamboo shoots and regulating soil phenolic acid after cultivation, characterized by, It comprises the following steps: (1) Cultivation of the pad: collect fresh, no disease and insect pests of sea mushroom dregs, after crushing mixed with urea, light calcium carbonate, according to 1.4-1.6 m high, 1.4-1.6 m wide, length unlimited size of stacking fermentation, every 10 days evenly turn over once, so that the sea mushroom dregs uniform fermentation, moisture content 60-65wt%, fermentation time 35-45 days; wherein the sea mushroom dregs, urea and light calcium carbonate mass ratio is 90:5:5; (2) Cultivation formula: the giant grass and sawdust according to one layer of giant grass and one layer of sawdust cycle stacking into 1.4-1.6 m high, 1.4-1.6 m wide, length unlimited size of stacking fermentation, every 10 days evenly turn over once, so that the sawdust and giant grass mixed uniform fermentation, moisture content 60-70wt%, fermentation time 40-45 days; wherein the giant grass and sawdust mass ratio is 1:1; (3) Application method: in the process of cultivation of bamboo corolla, according to the fermentation of sea mushroom dregs as the pad high 10 cm, the giant grass and sawdust after fermentation as the main culture material, according to the high 40 cm specifications for the laying of bamboo corolla, the bamboo corolla is divided into egg size after 30-50 cm each of the distance inoculation, and the soil cover is 3-5 cm thick, the bamboo corolla strain dosage is 200-230 bags / mu, 0.9 kg / bag, and the bamboo corolla mycelium recovery and fruiting body culture stage is carried out; (4) The soil conditioner is fermented by bacillus subtilis and fermented sea mushroom dregs, and the cultivated bamboo corolla soil is evenly treated from the third month before the next year of bamboo corolla cultivation, and the yield per mu of continuous cultivation in the second year basically recovers the same as the first year; wherein the dosage of bacillus subtilis in the soil conditioner is 1 kg / mu, the dosage of fermented sea mushroom dregs is 6 t / mu, and the soil is applied once before the cultivation of bamboo corolla in the third month.

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