Traditional Chinese medicine residue and mushroom dreg compound fermented fertilizer as well as preparation method and application thereof

Abstract

The invention relates to the technical field of fertilizer preparation, in particular to traditional Chinese medicine residue and mushroom dreg compound fermented fertilizer as well as a preparation method and application thereof. The preparation method of the traditional Chinese medicine residue and fungus residue compound fermented fertilizer comprises the following steps: (1) crushing pleurotus eryngii fungus residues, perilla leaf traditional Chinese medicine residues and cow dung, and mixing to obtain a material 1; and (2) adjusting the carbon nitrogen ratio and the water content of the material 1, and then fermenting. The fermented fertilizer provided by the invention can significantly influence the growth of potted tobacco, soil characteristics and microbial communities, and provides a theoretical basis and practical guidance for optimizing tobacco cultivation and fungus residue utilization.

Description

Technical Field

[0001] The invention relates to the technical field of fertilizer preparation, and in particular to a composite fermentation fertilizer of traditional Chinese medicine residue and fungus residue, and a preparation method and application thereof. Background Art

[0002] In the current agricultural development process, the importance of recycling agricultural production waste is becoming increasingly prominent. Traditional Chinese medicine residue is a substance rich in cellulose, hemicellulose, lignin and other components, with an annual output of more than 35 million tons. However, it has not yet been fully developed and utilized. The output of edible fungus residue is huge and rich in nutrients such as protein and polysaccharides, but the degree of resource utilization is low. If these traditional Chinese medicine residues and Pleurotus eryngii residues are not fully utilized, it will not only cause a huge waste of resources, but also pollute the surrounding environment if not properly handled.

[0003] In the past few decades, agricultural waste treatment and resource recycling have received widespread attention and have become one of the hot research areas. During this period, new treatment technologies and methods have emerged continuously. Among them, composting technology, as an effective way of resource recycling, has stood out among many technical methods and has been highly valued by academia and industry. This technology provides a valuable way to solve the problem of agricultural waste and achieve sustainable resource utilization. Many scholars have conducted composting-related research on various types of agricultural waste such as grape branches, pig manure, and sludge. By adding mixed microorganisms, nitrification inhibitors, and rice husk charcoal, the composting speed is effectively accelerated, thereby improving the quality of the compost product. At the same time, some studies focus on the separate composting of Chinese medicine residues and edible fungus residues. For the composting of Chinese medicine residues, the key is to promote the degradation of cellulose and lignin; while the composting of edible fungus residues needs to focus on the conversion process of nitrogen. However, the current research on the mixed composting fermentation of Chinese medicine residues and Pleurotus eryngii residues is still insufficient. There are still many unknown areas that need to be explored in terms of determining the appropriate ratio of the two in mixed composting, precise control of key parameters, and accurate judgment of the degree of maturity.

[0004] In the field of agricultural development, the composting and fermentation of traditional Chinese medicine residue and edible fungus residue to produce organic fertilizers shows broad prospects. Studies have shown that such organic fertilizers play a positive role in improving soil structure and enhancing soil fertility. In the composting and fermentation process, microorganisms are key factors, and their types, quantity and activity have a significant impact on fermentation efficiency and product quality that cannot be ignored. However, current research on the mixed fermentation of traditional Chinese medicine residue and edible fungus residue is still insufficient. In this mixed fermentation scenario, the characteristics of the microbial community and the specific impact of changes in the bacterial community are still unknown, which leaves a large number of research gaps in this field that need to be filled. Summary of the invention

[0005] The purpose of the present invention is to provide a composite fermentation fertilizer of Chinese medicine residue and mushroom residue. The present invention selects Chinese medicine residue of Perilla leaf, mushroom residue of Pleurotus eryngii and cow dung to mix and build a pile for natural fermentation, comprehensively monitors the changes in composting fermentation parameters and nutritional components, and deeply analyzes the changes in bacterial communities. The present invention has important theoretical significance for in-depth understanding of the mixed composting mechanism, and also provides an innovative method for efficient treatment of agricultural waste and production of organic fertilizers, and has high practical application value.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a method for preparing a composite fermentation fertilizer of traditional Chinese medicine residues and fungus residues, comprising the following steps:

[0008] (1) Pleurotus eryngii residue, Perilla leaf Chinese medicine residue and cow dung are crushed and mixed to obtain material 1;

[0009] (2) adjusting the carbon-nitrogen ratio and the water content of the material 1 and then fermenting it.

[0010] Preferably, in step (1), the mass ratio of the Pleurotus eryngii residue, the Perilla leaf Chinese medicinal residue and the cow dung is 8-12:0.5-1.5:0.5-1.5.

[0011] Preferably, the diameter of the material after crushing in step (1) is ≤0.5 cm.

[0012] Preferably, the carbon-nitrogen ratio in step (2) is adjusted to 23-27.

[0013] Preferably, the water content in step (2) is adjusted to 50-70%.

[0014] Preferably, the fermentation time in step (2) is 70 to 90 days, and the compost is turned every 8 to 12 days.

[0015] The invention also provides a composite fermented fertilizer of traditional Chinese medicine residue and fungus residue prepared by the preparation method.

[0016] The invention also provides application of the traditional Chinese medicine residue and fungus residue composite fermentation fertilizer in soil improvement.

[0017] The invention also provides application of the traditional Chinese medicine residue and fungus residue composite fermentation fertilizer in tobacco planting.

[0018] The invention also provides application of the traditional Chinese medicine residue and fungus residue composite fermentation fertilizer in cabbage planting.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The composite fermentation fertilizer of Chinese medicine residue and fungus residue prepared by the method of the present invention has a significant effect on the activities of soil dehydrogenase, catalase, polyphenol oxidase and nitrate reductase, and significantly improves the contents of soil organic matter, alkaline nitrogen, acidic available phosphorus and available potassium; the structure and diversity of soil fungal communities change significantly under different addition amounts, different addition amounts have different dominant microbial communities, and the relationship between different dominant fungal genera and enzyme activities and soil nutrients is different. The agronomic traits of tobacco plant height, leaf number and other growth, different addition amounts at different times have different promotion effects; the nicotine content in tobacco leaves increases with the increase of substrate, the reducing sugar content decreases, the chlorine content decreases, and the potassium content is stable; the nitrogen and chlorophyll content of the lower leaves is significantly increased, the middle leaves are relatively stable, and the upper leaves are less affected. It shows that the fermentation substrate of fungus residue can significantly affect the growth of potted tobacco, soil characteristics and microbial communities, and provides a theoretical basis and practical guidance for optimizing tobacco cultivation and the utilization of fungus residue. In addition, the fertilizer provided by the present invention also has the effect of improving the germination index of cabbage, and has a significant growth-promoting effect on cabbage. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The following is an analysis chart of the dynamic changes of temperature, humidity, pH value and conductivity during the composting process in Example 1; A: temperature change; B: moisture content change; C: pH value change; D: EC value change.

[0022] Figure 2 This is an analysis chart of changes in physical properties of materials during composting fermentation in Example 1; A: change in bulk density; B: change in water holding porosity; C: change in total porosity; D: change in ventilation porosity.

[0023] Figure 3 The nutrient, organic matter and germination index analysis diagram of the composting fermentation process in Example 1; A: change in alkaline nitrogen content; B: change in available phosphorus content; C: change in available potassium content; D: change in humus content; E: change in organic matter content; F: change in germination index.

[0024] Figure 4 This is an analysis diagram of the evolution of bacterial community structure, diversity changes and correlation with environmental factors during the composting process of Example 1; A: species composition; B: bacterial community structure; C: Chao1 index; D: Simpson index; E: Shannon index; F: principal component analysis; G: dominant bacterial group; H: Spearman correlation.

[0025] Figure 5 This is Example 2, the effect of compost on enzyme activity in potted tobacco soil.

[0026] Figure 6 This is the effect of compost on the nutrient content of potted tobacco soil in Example 2.

[0027] Figure 7This is Example 2, the effect of composting on the fungal community in the soil of potted tobacco.

[0028] Figure 8 This is a graph showing the content of dominant microorganisms and other types of microorganisms in Example 2.

[0029] Fig. 9 This is the effect of compost on the growth and development of potted tobacco in Example 2.

[0030] Fig.10 This is the effect of composting on the relative contents of N and chlorophyll in different parts of potted tobacco in Example 2.

[0031] Fig.11 This is Example 2, the effect of compost on the chemical composition of potted tobacco leaves.

[0032] Fig.12 The effects of each treatment in Example 3 on the agronomic traits of flue-cured tobacco are shown in FIG.

[0033] Fig.13 The effects of each treatment in Example 3 on the increase and growth rate of agronomic traits of flue-cured tobacco. DETAILED DESCRIPTION

[0034] The present invention provides a method for preparing a composite fermentation fertilizer of traditional Chinese medicine residues and fungus residues, comprising the following steps:

[0035] (1) Pleurotus eryngii residue, Perilla leaf Chinese medicine residue and cow dung are crushed and mixed to obtain material 1;

[0036] (2) adjusting the carbon-nitrogen ratio and the water content of the material 1 and then fermenting it.

[0037] In the present invention, the mass ratio of the Pleurotus eryngii residue, the Perilla leaf Chinese medicinal residue and the cow dung in step (1) is 8-12:0.5-1.5:0.5-1.5; preferably 9-11:0.7-1.3:0.7-1.3; further preferably 10:0.9-1.1:0.9-1.1; more preferably 10:1:1.

[0038] In the present invention, the diameter of the material after the crushing in step (1) is ≤0.5 cm; preferably 0.3-0.5 cm; more preferably 0.4 cm.

[0039] In the present invention, the carbon-nitrogen ratio in step (2) is adjusted to 23-27; preferably 24-26; and more preferably 25.

[0040] In the present invention, the water content in step (2) is adjusted to 50-70%, preferably 54-66%, more preferably 58-62%, and more preferably 60%.

[0041] In the present invention, the fermentation time of step (2) is 70 to 90 days, and the compost is turned every 8 to 12 days; preferably, the fermentation time is 74 to 86 days, and the compost is turned every 9 to 11 days; further preferably, the fermentation time is 78 to 82 days, and the compost is turned every 10 days; more preferably, the fermentation time is 80 days, and the compost is turned every 10 days.

[0042] The invention also provides a composite fermented fertilizer of traditional Chinese medicine residue and fungus residue prepared by the preparation method.

[0043] The invention also provides application of the traditional Chinese medicine residue and fungus residue composite fermentation fertilizer in soil improvement.

[0044] The invention also provides application of the traditional Chinese medicine residue and fungus residue composite fermentation fertilizer in tobacco planting.

[0045] The invention also provides application of the traditional Chinese medicine residue and fungus residue composite fermentation fertilizer in cabbage planting.

[0046] In the present invention, the Pleurotus eryngii residue is provided by Hunan Yuxiu Biotechnology Co., Ltd., the Perilla leaf Chinese medicine residue comes from Hunan Times Sunshine Pharmaceutical Co., Ltd., and the cow dung is supplied by Yongzhou Siyuan Agricultural Technology Co., Ltd.

[0047] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0048] Example 1

[0049] 1.1 Experimental Materials The experiment was carried out at the Tobacco Leaf Production Technology Center of Yongzhou Tobacco Company from July to October 2023. The starting material was 100t. The material was first crushed to a particle diameter of less than 0.5cm, and then mixed with Pleurotus eryngii residue, Perilla leaf Chinese medicine residue and cow dung in a mass ratio of 10:1:1 by a large mixer. The carbon-nitrogen ratio was adjusted to 25 with urea, and the moisture content was sprayed to 60%. Three piles (15m×3m×1.5m) were piled for natural fermentation, and the piles were turned every 10d for a total of 80d, with the first day counted as the first day. A multi-channel temperature recorder was used to record the temperature, moisture content, pH value and EC value of the pile every day; samples were collected from the top, middle and bottom of the pile by the diagonal five-point sampling method during the heating period (1st day), high temperature period (5th day), cooling period (42nd day) and maturity period (61st day), and divided into two parts after sufficient mixing. The first part was air-dried at room temperature, crushed and sieved through a 0.1 mm sieve for the determination of physical properties, organic matter, nitrogen, phosphorus, potassium and humus; the second part was stored at -80°C to evaluate bacterial diversity.

[0050] 1.2 Determination indicators and methods

[0051] (1) Physical and chemical indicators: Organic matter, nitrogen, phosphorus and potassium were determined according to the standard methods of "Organic Fertilizer" (NY / T 525-2021) and "Determination of Nitrate Nitrogen, Ammonium Nitrogen and Amide Nitrogen Content in Fertilizers" (NY / T 1116-2014); bulk density, total porosity, aeration porosity, water holding porosity and humus were determined according to the literature [Bao Shidan. Soil Agrochemical Analysis (3rd Edition) [M]. Beijing: China Agriculture Press, 2000.].

[0052] (2) Seed germination index: Weigh 10 g of sample, add 50 mL of distilled water, shake on a shaker for 30 minutes, and centrifuge for 10 minutes to obtain the extract. Place filter paper in a sterile 9 cm culture dish, take 20 uniform and full cabbage seeds and put them into the dish, add 5 mL of extract, use distilled water as a control, and culture in a constant temperature culture room at 25°C for 72 hours. Measure the germination rate and root length. Repeat each treatment (including the control) three times.

[0053] Germination index GI (%) = (average germination rate of treatment × average root length of treatment) / (average germination rate of control × average root length of control) × 100.

[0054] (3) Bacterial community composition: 16S rRNA gene sequencing analysis was performed. The total bacterial DNA of the samples was extracted using the magnetic bead soil and feces genomic DNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.), the V3-V4 region of the 16S rRNA gene was amplified using primers 341F and 806R, the PCR products were sequenced using the Illumina platform, and microbial biological information analysis was performed using the Microscience Alliance bioinformatics analysis cloud platform.

[0055] 1.3 Data processing

[0056] Microsoft Excel 2019 was used to process and analyze the data, and variance analysis was performed with the help of SPSS22.0 (p<0.05 was considered a significant difference). The data were expressed as mean ± standard deviation.

[0057] 2 Results and analysis

[0058] 2.1 Analysis of dynamic changes of temperature, humidity, pH value and conductivity during composting

[0059] Depend on Figure 1 It can be seen that the entire fermentation process can be divided into four stages according to the temperature: the temperature rise period (1-4 days), the high temperature period (5-41 days), the temperature drop period (42-60 days) and the maturity period (61-80 days). Figure 1 -A). The moisture content is stable at about 62% during the temperature rise period, gradually decreases during the high temperature period, continues to slowly decrease during the temperature drop period, and basically stabilizes at a lower level during the maturity period ( Figure 1-B). The pH value is stable at 7.3-7.5 during the warming period, fluctuates and rises during the high temperature period, is relatively stable with small fluctuations during the cooling period, and is stable at 8.1-8.3 during the mature period ( Figure 1 -C). The conductivity rises slowly from 2.4 to 2.6 during the warming period, rises sharply to 4.9 during the high temperature period, rises slowly to 5.1 during the cooling period, and fluctuates between 4.8 and 5.3 during the mature period ( Figure 1 -D). This shows that at different fermentation stages, temperature, moisture content, pH value and conductivity all present different trends, reflecting the combined effects of multiple factors such as microbial activity and material transformation during the fermentation process.

[0060] 2.2 Analysis of changes in physical properties of materials during composting

[0061] Depend on Figure 2 It can be seen that during the whole process of composting, the bulk density and water holding porosity increased significantly, while the total porosity and ventilation porosity gradually decreased. In terms of bulk density, it was 0.26g / cm3 during the warming period, rose to 0.31g / cm3 during the high temperature period, reached 0.33g / cm3 during the cooling period, and stabilized at 0.34g / cm3 during the mature period ( Figure 2 -A); the water-holding porosity is 40.1% in the warming period, rises to 44.2% in the high temperature period, reaches 45.6% in the cooling period, and reaches as high as 46.2% in the mature period ( Figure 2 -B). For the total porosity, it was 71.3% during the temperature rise period, 68.1% during the high temperature period, 67.4% during the temperature drop period, and further decreased to 66.2% during the mature period ( Figure 2 -C); the ventilation porosity is 31.2% in the warming period, drops to 25.1% in the high temperature period, becomes 23.3% in the cooling period, and drops to 20.5% in the mature period ( Figure 2 -D). This indicates that as the fermentation progresses, the material gradually becomes denser and its water holding capacity increases, but its aeration performance weakens. This trend reflects the dynamic changes in the physical properties of the material during the composting fermentation process.

[0062] 2.3 Analysis of nutrients, organic matter and germination index during composting fermentation

[0063] Depend on Figure 3 It can be seen that the contents of alkaline hydrolyzed nitrogen, available phosphorus, quick-acting potassium and humus showed an upward trend in the four stages of fermentation. The alkaline hydrolyzed nitrogen content was 2.23g / kg in the warming period, increased to 2.87g / kg in the high temperature period, slightly decreased to 2.82g / kg in the cooling period, and reached 2.75g / kg in the mature period ( Figure 3 -A); the available phosphorus is 0.291g / kg during the warming period, rises to 0.323g / kg during the high temperature period, continues to rise to 0.338g / kg during the cooling period, and stabilizes at 0.342g / kg during the mature period ( Figure 3-B); the content of quick-acting potassium is 0.72g / kg in the rising temperature period, rises to 0.80g / kg in the high temperature period, continues to rise to 0.82g / kg in the falling temperature period, and reaches 0.85g / kg in the mature period ( Figure 3 -C); humus was 302.5g / kg in the rising temperature period, increased to 326.2g / kg in the high temperature period, increased to 336.4g / kg in the falling temperature period, and increased to 344.6g / kg in the mature period ( Figure 3 -D). This indicates that as fermentation progresses, nutrients are continuously transformed and accumulated, providing potential nutritional support for subsequent soil improvement and crop growth. However, the organic matter content decreased, from 442.7 g / kg during the warming period to 419.2 g / kg during the high temperature period, slowly decreasing to 410.4 g / kg during the cooling period, and continuously decreasing to 404.2 g / kg during the mature period ( Figure 3 -E), which reflects the decomposition and transformation of organic matter during the fermentation process. The germination index first dropped and then rose, from 62.3% in the warming period to 52.1% in the high temperature period, to 73.2% in the cooling period, and continued to rise to 93.5% in the mature period ( Figure 3 -F), indicating that there may be some factors that are not conducive to seed germination in the early stage of fermentation, but as the fermentation proceeds, the compost products gradually mature, the inhibitory effect on seed germination weakens, and even turns into a promoting effect.

[0064] 2.4 Analysis of the evolution of composting bacterial community structure, diversity changes and correlation with environmental factors

[0065] Species composition analysis ( Figure 4 A) shows that the kingdom level is consistent in fermentation, and the value increases when transitioning to the species level, which means that the bacterial community has evolved from the initial composition to a more diverse and refined structure. Although the bacterial community structure varies greatly at each stage, there are 14 core bacterial groups ( Figure 4 B). Chao1 index rises ( Figure 4 C), indicating that the richness of the bacterial community increased. The Simpson and Shannon indices were low during the warming period and increased in other stages, indicating that as the fermentation progressed, the diversity and uniformity of the bacterial community increased and the dominant position of the dominant bacterial community weakened ( Figure 4 DE). Different stages have different distributions on Axis1, Axis2, and Axis3 of PCoA ( Figure 4 F), indicating that fermentation changes the bacterial community structure and increases with the stage. Firmicutes dominate the warming period, Proteobacteria account for a high proportion in the high temperature period, Firmicutes and Thermococcus dominate in the cooling period, and Actinobacteria, Firmicutes and Bacteroidetes dominate in the mature period ( Figure 4G). Spearman correlation analysis showed that Chloroflexi, Actinobacteria, and Firmicutes_G were negatively correlated with water, and Proteobacteria were positively correlated; Proteobacteria and Firmicutes_D were negatively correlated with conductivity, and Actinobacteria, Chloroflexi, and Firmicutes_G were positively correlated; Bacteroidetes were negatively correlated with pH ( Figure 4 H). This indicates that environmental factors in compost fermentation affect the growth and metabolism of dominant bacterial phyla, thereby affecting the bacterial community structure, function, fermentation efficiency and quality.

[0066] The results of this study show that composting is divided into four stages: heating, high temperature, cooling, and maturity. During this process, the relevant parameters, physical properties, nutrient content, and bacterial communities show regular changes. This is consistent with the results of previous studies. However, our results differ from those of Ma et al. (Ma Y, Liu L, Zhou X, Tian T, Xu S, Li D, Li C, Li Y. Optimizing Straw-Rotting Cultivation for Sustainable Edible Mushroom Production: Composting Spent Mushroom Substrate with Straw Additions. J Fungi (Basel). 2023 Sep 13; 9 (9): 925.). This may be due to the different chemical composition of the compost raw materials, which leads to different degradation characteristics. In addition, the addition of bacterial preparations by Ma et al. changed the core microbial flora, causing it to heat up faster, but the high temperature period lasted for a shorter period of time. In this study, we found that as the fermentation progressed, the bacterial community structure became richer and more complex, and the original dominant position of the dominant flora also changed.

[0067] Further in-depth study of bacterial changes during the composting process revealed that different stages had different dominant bacterial communities. Firmicutes dominated the warming period. During this period, thermophilic bacteria degraded organic matter to produce small molecules and heat, which increased the temperature. The growth pattern of Firmicutes in the warming period of different composting systems may be universal. Based on this, in the warming period, we can promote the growth of Firmicutes according to their needs through reasonable material selection and pretreatment, such as increasing the content of easily decomposable organic matter, and thus promote the composting temperature. When entering the high temperature period, thermophilic microorganisms become the dominant bacterial community, they continue to degrade organic matter and begin to form humus. At this stage, Proteobacteria accounted for a high proportion, which highlights its key role in accelerating the decomposition of organic matter and the initial formation of humus in the high temperature composting stage. As the composting process enters the cooling period, the activity of thermophilic microorganisms decreases, most of the organic matter has been degraded, and the compost temperature drops. At this time, Firmicutes become the dominant bacteria, which reflects the importance of Firmicutes in the multi-stage composting process, which can ensure a smooth transition of the composting process. In the mature stage, mesophilic microorganisms become active again, degrading refractory substances, increasing the degree of humification and stabilizing it. At this stage, Actinomycetes, Firmicutes and Bacteroidetes become the main dominant bacteria, and these three together promote the maturity of compost. The synergistic effect between them is crucial to the quality and maturity of compost.

[0068] Example 2

[0069] 1.1 Test materials

[0070] The test soil was taken from the 0-20 cm tillage layer of the tobacco test base of the Tobacco Leaf Production Technology Center of Yongzhou City. It was red soil with an alkaline nitrogen content of 199.0 mg / kg and a pH of 6.05. The test material was "Yunyan 87".

[0071] Preparation of fungus and medicinal residue fermentation matrix: prepared according to the method in Example 1.

[0072] 1.2 Experimental design After the fermentation matrix of fungus and drug residues was fully mixed with the test soil, it was filled into a plastic pot with a length of 40 cm, a width of 38 cm, and a height of 30 cm. This experiment set up four treatments (see Table 1 for details), each treatment set up 3 replicates, and each replicate contained 5 pots. On March 29, 2024, one "Yunyan 87" tobacco plant (sown on December 27) was transplanted in each pot, and all pots were placed in the sun room of the tobacco test base of Yongzhou Tobacco Leaf Production Technology Center, and arranged in a randomized block arrangement. No fertilizer was applied during the entire experimental period. On April 29, 2024, the agronomic traits of tobacco plants were measured. On May 31, 2024, the agronomic traits of tobacco plants were measured, and the relative contents of N and chlorophyll in tobacco leaves were measured; at the same time, the largest leaf on the upper part of the tobacco plant was collected, and its chemical composition was detected after being processed according to the standard baking process of Yongzhou Tobacco Company of Hunan Tobacco Company. In addition, 5 pots were randomly selected for each treatment, and 60 g of soil from the 10-20 cm soil layer was collected in each pot using the five-point sampling method. The collected soil was mixed evenly to form a sample. Subsequently, each sample was divided into two parts. The first part was air-dried at room temperature and crushed, and passed through a 0.1 mm sieve for the determination of soil physical and chemical indicators; the second part was stored in an environment of -80 ° C for the evaluation of soil fungal community composition.

[0073] Table 1 Edible fungus residue dosage

[0074]

[0075] 1.3 Determination indicators and methods

[0076] (1) Physical and chemical indicators: Soil organic matter, alkaline nitrogen, acidic available phosphorus and available potassium contents were determined according to the literature [Bao Shidan. Soil Agrochemical Analysis (3rd Edition) [M]. Beijing: China Agriculture Press, 2000.].

[0077] (2) Agronomic traits: maximum leaf length, maximum leaf width, plant height, stem girth, maximum leaf weight, number of leaves, internode distance, etc. The measurement standards of various agronomic traits are implemented in accordance with the national standard YC / T142-1998 [State Tobacco Monopoly Administration, 1998].

[0078] (3) Leaf chemical composition: Reducing sugar, nicotine, and chlorine were detected by continuous flow method, and potassium was detected by flame photometry [Wang Xichun, Yuan Fang, Zhou Xiangping, et al. Effects of different nitrogen application levels on agronomic traits and product quality of 'Xiangyan No. 5' [J]. Chinese Agricultural Science Bulletin, 2018, 34(26): 31-36.].

[0079] (4) Relative contents of nitrogen and chlorophyll: detected using Zhejiang Top TYS-4N plant nutrition analyzer.

[0080] (5) Fungal community composition: The total microbial DNA of the samples was extracted using a magnetic bead soil and feces genomic DNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.). The 18S rRNA was amplified using primers ITS1F and ITS2R. The PCR products were sequenced using the Illumina platform, and the microbial biological information analysis was performed using the Microscience Alliance bioinformatics analysis cloud platform.

[0081] 1.5 Data Statistical Analysis

[0082] Microsoft Excel 2019 was used to process and analyze the data, and variance analysis was performed with the help of SPSS22.0 (p<0.05 was considered a significant difference). The data were expressed as mean ± standard deviation.

[0083] 2 Results and analysis

[0084] 2.1 Effects on enzyme activities in potted tobacco soil

[0085] As the amount of fermentation substrate added to the fungus residue increased from 0 to 15%, the activities of dehydrogenase, catalase, polyphenol oxidase and nitrate reductase showed varying degrees of change ( Figure 5 The dehydrogenase activity gradually increased and significantly increased when the addition amount was 15% ( Figure 5 A); Catalase activity continued to rise and reached a maximum value when the addition amount reached 15% ( Figure 5 B); the activity of polyphenol oxidase increased slightly when the addition amount was from 0 to 5% and 10%, and increased significantly when the addition amount was 15% ( Figure 5 C); the activity of nitrate reductase also increased, especially when the addition amount was 15% ( Figure 5 D), indicating that the addition amount of fermentation substrate of fungus residue has a significant effect on the activities of these enzymes in the soil, and the response degree of different enzymes to the addition amount varies, reflecting the close relationship between substrate addition and soil enzyme activity.

[0086] 2.2 Effects on soil nutrient content of potted tobacco

[0087] With the increase of the amount of fermented substrate added, the contents of soil organic matter, alkaline nitrogen, acidic available phosphorus and available potassium all showed a significant upward trend ( Figure 6 Among them, the soil organic matter content gradually increased from 22.7g / kg to 53.6g / kg, 64.0g / kg, and 105.3g / kg ( Figure 6 A); the alkaline nitrogen content increased from 198.9mg / kg to 360.4mg / kg, 417.1mg / kg, 695.6mg / kg ( Figure 6B); the acidic available phosphorus content gradually increased from 22.9 mg / kg to 174.2 mg / kg, 232.6 mg / kg, and 395.6 mg / kg ( Figure 6 C); the effective potassium content increased steadily from 95.3 mg / kg to 350.1 mg / kg, 522.6 mg / kg, 1116.1 mg / kg ( Figure 6 D). This fully demonstrates that the fermented matrix of fungus and medicinal residue has a significant effect on improving soil nutrients and can be used as a soil conditioner to improve soil fertility.

[0088] 2.3 Effects on soil fungal communities in potted tobacco

[0089] The number of fungi at each taxonomic level from kingdom to species in each treatment soil showed different trends. The number of microorganisms at each taxonomic level was low at 0% addition, increased significantly at 5%, higher than 0% but lower than 5% at 10%, and the lowest at 15% ( Figure 7 A). The number of unique OTUs in each treatment was 717, 1281, 1113 and 621 respectively, and there was no common OTU, indicating that each treatment formed a unique fungal community structure in the soil, lacking a common core fungal group ( Figure 7 B). Chao1 index showed that the T1 group significantly increased the richness of microbial species, followed by the CK group, the T2 group had a certain reduction effect, and the T3 group strongly inhibited the species richness ( Figure 7 C); Shannon index showed that the T1 group significantly improved the diversity of microbial communities, followed by the CK group, the T2 group had no significant effect, and the T3 group significantly inhibited the diversity ( Figure 7 D). PCoA analysis showed that the fungal community structure of the CK group had a relatively stable distribution range in three-dimensional space; as the amount of addition increased, the distribution of the microbial community structure gradually moved away from the CK group ( Figure 7 E), suggesting that low additions cause smaller changes, while high additions lead to larger adjustments. The dominant fungal communities in the soil of the CK group are Torula, Lecanicillium, Monilia, and Cercophora; T1 is Penicillium, Iodophanus, and Saitozyma; T2 is Leucocoprinus, Aspergillus, Fusarium, and Arthrobotrys; and T3 is Metacordyceps, Cladosporium, Monocilllium, and Trichoderma. This shows that each treatment creates a unique soil ecological environment, and those that adapt to the conditions become the dominant microorganisms ( Figure 8 The relationship between different dominant fungal genera and enzyme activity and soil nutrients varies. Leucocoprinus, Cladosporium, Metacordyceps, and Monocillium have a positive effect on improving enzyme activity and soil nutrients, while Clydaea, Rhizophagus, Conocybe, Sphaerobolus, Torula, Lecanicillium, and Cercophora have the opposite effect. The others have relatively small effects ( Figure 8 below).

[0090] 2.3 Effects on the growth and development of potted tobacco

[0091] At 31d and 61d, the plant height, leaf number, stem girth, leaf length and leaf width of all tobacco treatments showed an increasing trend ( Fig. 9 ), reflecting the normal growth process of tobacco. At 31 days, all tobacco plants grew normally ( Fig. 9 A), different addition amounts have different effects on growth indicators. In terms of plant height, T1 and T2 are outstanding. In terms of leaf number, T1 and T2 are relatively better. In terms of stem girth, T2 has an advantage. In terms of leaf length and leaf width, T1 has a larger value, followed by T2 ( Fig. 9 B). At 61 days, the bottom leaves turned yellow, and the condition gradually improved with the increase of the dosage ( Fig. 9 C), its growth promoting effect was more significant than that of 31d, and its 15% addition had a particularly prominent promoting effect on most indicators. This indicates that the different addition amounts of fungus residue fermentation substrates have a time- and dose-dependent effect on agronomic traits during tobacco growth.

[0092] 2.4 Effects on N content and relative chlorophyll content in different parts of potted tobacco

[0093] Different addition amounts of fermented substrates from fungus and medicinal residues have different effects on the nitrogen content and relative content of chlorophyll in tobacco leaves from different parts ( Fig.10 ). Lower leaves: Nitrogen content increased from 0.8mg / g to 1.6mg / g, and chlorophyll content increased from 3.0 to 15.6; 5%, 10%, and 15% additions compared with the control showed significant differences in these two indicators, indicating that it can promote the increase of nitrogen content and chlorophyll synthesis in the lower leaves; Middle leaves: Nitrogen content increased from 1.8mg / g to 2.6mg / g, and chlorophyll content increased from 17.5 to 29.6. Within the range of 5% to 15%, there was no significant difference between the two indicators, indicating that this range has a stable effect on the improvement of middle leaves, and increasing the addition has no obvious effect; Upper leaves: Nitrogen content is stable at around 3.2mg / g, and chlorophyll content is stable at around 37.0. There is no significant difference between the different addition treatments, and the effect on the upper leaves is small ( Fig.10A and B). This shows that the effects of different amounts of added bacterial residue fermentation substrate on the nitrogen content and relative chlorophyll content of tobacco leaves in different parts have obvious site specificity.

[0094] 2.5 Effects on the chemical composition of potted tobacco leaves

[0095] The effects of different amounts of added fungus residue fermentation substrate on the contents of nicotine, reducing sugar, chlorine and potassium in potted tobacco leaves were different ( Fig.11 The nicotine content was 2.5g / kg when the addition amount was 0%, and it increased to 2.7g / kg, 3.0g / kg, and 3.6g / kg when the addition amount increased to 5%, 10%, and 15%, respectively. Fig.11 A), which may promote the synthesis of nicotine. The reducing sugar content was 14.2 when the addition amount was 0, and when the addition amount increased to 5%, 10%, and 15%, it decreased to 12.9, 12.4, and 9.7 ( Fig.11 B), indicating that it has an inhibitory effect on the accumulation of reducing sugars. The chlorine content was 1.2 when the addition amount was 0, and dropped to 0.7, 0.6, and 0.8 ( Fig.11 C), indicating that it can reduce the chlorine content in tobacco leaves. There is no obvious effect on potassium content, which is stable at around 1.3, with no significant difference ( Fig.11 D). It shows that different addition amounts of fermented substrates of fungus and medicinal residues have different effects on the contents of nicotine, reducing sugar, chlorine and potassium, the main chemical components in potted tobacco leaves.

[0096] It can be seen that fungus residue is rich in a large number of nutrients such as nitrogen, phosphorus, and potassium. After returning to the field, it can increase soil nutrients, improve soil structure, and enhance soil fertility. With the increase in the amount of fermentation matrix added by fungus residue, the activity of enzymes in the soil showed different degrees of change; at the same time, the fermentation matrix of fungus residue significantly increased the nutrient content of the soil and changed the structure and diversity of soil microbial communities. Studies have shown that returning fungus residue to the field can increase the soil enzyme activity of paddy field soil, increase soil nutrients, and significantly increase the number of soil microorganisms. Further studies have found that different addition amounts have different dominant microbial communities, some of which have a positive effect on enzyme activity and soil nutrients, while others have the opposite effect. The fermentation matrix of fungus residue contains rich nutrients, and some dominant microbial communities may affect the transformation and effectiveness of nutrients in the soil. For example, Cladosporium can decompose organic matter in the soil into simple inorganic forms, thereby improving the effectiveness of nutrients in the soil. The reason why different addition amounts lead to changes in microbial communities may be related to the number of nutrients available to microorganisms in the soil. As the amount of addition increases, the concentration of nutrients in the matrix changes, which may provide different living conditions for different microorganisms, leading to changes in the structure of the microbial community.

[0097] Example 3

[0098] 1.1 Test materials

[0099] The flue-cured tobacco variety selected is Xiangyan No. 7. Within the tobacco planting area of ​​Yongzhou Base of Central South Station, the selected test field has not carried out fertilizer test-related activities in the past three years. The field fertility of the test field is at a medium level, and the fertility distribution shows a consistent characteristic. At the same time, it has good drainage and irrigation performance, which can provide relatively stable and suitable soil environment conditions for flue-cured tobacco growth, and ensure the stability and controllability of the influence of soil fertility factors on flue-cured tobacco growth during the test, so as to more accurately explore the influence of other factors on flue-cured tobacco growth and related indicators.

[0100] 1.2 Experimental setup

[0101] The experiment was carried out in 2024 at the Tobacco Experimental Base of Yongzhou Tobacco Production Technology Center. A single-factor completely randomized block design was used, with 4 treatments (T1, T2, T3 and T4) (Table 2), and each treatment was repeated 3 times for a total of 12 plots (Table 3). 100 flue-cured tobacco plants were planted in each plot, with a row spacing of 1.2m and a plant spacing of 0.5m. Before cultivation, the fermentation matrix of the fungus and drug residue (prepared by the method in Example 1) was sprinkled on the ridges according to the dosage and turned into the soil to mix. Two or more protective rows were set up around to reduce interference and ensure environmental stability. Field cultivation management was carried out in accordance with the Yongzhou Tobacco Production Technical Plan to ensure scientific and consistent experimental conditions.

[0102] Table 2 Matrix dosage

[0103]

[0104] Table 3 Field design

[0105]

[0106] 1.3 Fertilization method

[0107] The basal fertilizer adopts the "101" fertilization method: after completing the land preparation and ridge-forming operations, dig planting holes on the tobacco ridges at a spacing of 0.5 meters. Before transplanting the flue-cured tobacco, apply 30% of the total basal fertilizer (including special basal fertilizer and phosphate fertilizer) into the planting holes, and mix the basal fertilizer with the soil thoroughly and evenly. This is the "0" fertilizer part. Before or after transplanting the flue-cured tobacco, open a fertilization belt with a length of 15 cm and a depth of 15 cm between the two planting holes along the direction perpendicular to the tobacco ridge, and apply 70% of the total basal fertilizer, which is the "1" fertilizer.

[0108] Topdressing method: Take 2kg and 3kg of seedling fertilizer at the time of transplanting and 7 days after transplanting, respectively, and add water for watering. About 15 days after transplanting, apply half of the dosage of special topdressing. About 35 days after transplanting (before the clustering stage), apply the remaining special topdressing. Apply the remaining potassium sulfate before 45 days after transplanting. In rainy weather conditions, topdressing is suitable for dry application by hole digging; in dry weather conditions, it is appropriate to apply by hole digging, and the hole digging depth should be controlled in the range of 15-20cm.

[0109] This fertilization plan is designed to reasonably supply nutrients according to different weather conditions and tobacco growth stages to ensure the nutritional needs of tobacco during its growth process and promote its good growth and development. At the same time, it also fully considers the impact of environmental factors in actual operations on the fertilization effect, and improves fertilizer utilization through scientific fertilization methods to provide guarantees for the yield and quality of flue-cured tobacco.

[0110] 1.4 Field Management The test field adopts the bare land planting mode, the transplanting period is set as March 19, and the transplanting density is set at 1.2 meters × 0.5 meters. For field management methods not explicitly mentioned, they shall be strictly implemented in accordance with the "2024 Yongzhou Flue-cured Tobacco Production Technical Plan". Throughout the test process, it is imperative to ensure a high degree of consistency in agricultural operations in each plot, and minimize the experimental errors that may be caused by human factors through fine operating specifications and strict implementation standards.

[0111] 1.5 Survey indicators for each treatment 1.5.1 Survey methods for each treatment during the growth period

[0112] Twenty representative tobacco plants were selected from each treatment area as fixed-point observation objects, and the following growth periods were carefully observed and recorded: sowing period, which is the time point when sowing of flue-cured tobacco seeds began; transplanting period, which refers to the specific date when the tobacco seedlings were transplanted to the field; clumping period, during which the leaves of the tobacco plants began to lean against each other and form a clumping shape; vigorous growth period, when the tobacco plants grew rapidly and the stems and leaves grew vigorously; topping period, when the tobacco plants were toppled to control the apical growth and promote the distribution of nutrients to the leaves and other parts; maturity period, when the tobacco leaves reached maturity and could be harvested; field growth period, which is the time span of the entire field growth stage from transplanting period to maturity period.

[0113] 1.5.2 Investigation methods of agronomic traits of each flue-cured tobacco treatment

[0114] Ten representative tobacco plants were selected from the central area of ​​each plot and set as fixed-point observation plants. At 40 days, 55 days and 75 days after transplanting, the agronomic traits of the lower leaves, middle leaves and upper leaves were investigated and recorded in detail. The specific survey contents include: plant height, that is, the vertical height of the tobacco plant from the ground to the top growth point; number of leaves, the number of leaves of the tobacco plant; stem girth, the circumference measured at a specific position of the tobacco plant stem; maximum leaf length and width, respectively measuring the corresponding dimensions of the leaves with the largest length and width among the tobacco plant leaves; maximum leaf area, calculated by the formula "leaf area = length × width × 0.6345".

[0115] 1.5.3 Economic characteristics survey and sampling methods

[0116] When the tobacco leaves reach maturity and are ready for harvest, the tobacco leaves in each plot are separately hung and braided. Subsequently, the tobacco leaves in each plot are placed under uniform conditions for baking. After baking, the yield, output value, average price and tobacco grade ratio of each plot of baked tobacco are carefully investigated and analyzed. At the same time, C3F and B2F grade tobacco leaves are selected as sample tobacco to further test and study their relevant quality characteristics.

[0117] 2 Experimental results and analysis 2.1 Effects of each treatment on the growth period of flue-cured tobacco

[0118] As the addition of the fermented substrate of fungus and medicinal residue increased from 0% to 15%, the clustering period, vigorous growth period, budding period, topping period and flue-cured period all changed to varying degrees (Table 4). The clustering period was 37 days when the addition amount was 0%, and 31 days when the addition amount was 5%, 10% and 15%, indicating that it can accelerate the arrival of the clustering period and promote the early growth of flue-cured tobacco. The vigorous growth period was 42 days when the addition amount was 0%, and 36 days, 36 days and 34 days when the addition amount was 5%, 10% and 15%, respectively, which helped to advance the vigorous growth period and promote growth and material accumulation. The budding period was 67 days when the addition amount was 0%, 60 days when the addition amount was 5% and 10%, and 62 days when the addition amount was 15%. The overall situation first decreased and then slightly rebounded. It was advanced when the addition amount was 5% and 10%, and slightly extended when the addition amount was 15%, which may be affected by a combination of factors. The topping period was 74 days when the addition amount was 0%, and 67 days when the addition amount was 5%, 10%, and 15%, which was significantly advanced, which was related to growth rate and nutrient distribution, and may affect yield and quality. The harvesting and curing period was 77 days when the addition amount was 0%, and 70 days when the addition amount was 5%, 10%, and 15%, which was beneficial to production arrangement and processing in advance, and it was necessary to combine the analysis of tobacco leaf quality indicators. The killing period was 137 days at all addition amounts, which was not affected by the addition amount of fungus and drug residue fermentation matrix, and may be affected by other specific factors such as natural cycles. In summary, different addition amounts of fungus and drug residue fermentation matrix have a significant effect on the growth period of flue-cured tobacco, which is mainly reflected in the advance or change of the clustering period, vigorous period, budding period, topping period, and harvesting and curing period. These changes are of great significance to the growth and development and production management of flue-cured tobacco, and provide a reference for optimizing the application of fungus and drug residue fermentation matrix in flue-cured tobacco cultivation.

[0119] Table 4 Effects on the growth period of flue-cured tobacco in the field (d)

[0120]

[0121] 2.2 Effects of each treatment on agronomic traits of flue-cured tobacco

[0122] From the cluster stage to the vigorous growth stage and then to the dome stage, with the increase of the amount of fermented substrate added, the plant height, leaf number, stem girth, maximum leaf length, maximum leaf width and maximum leaf area of ​​flue-cured tobacco showed a gradual increase overall. This shows that the fermented substrate of fungus and medicinal residue has a promoting effect on the growth and development of flue-cured tobacco, and it is reflected in all growth stages and various indicators ( Fig.12 ; A plant height; B number of leaves; C stem girth; D leaf length; E leaf width; F leaf area).

[0123] From the cluster stage to the vigorous growth stage, the plant height growth rate of 0% addition was the smallest (32.8), which was significantly different from other additions, and the other additions were close in magnitude; from the vigorous growth stage to the round-top stage, the plant height growth rate of 0% addition was the largest (65.7cm), which was significantly different from other additions, and the other additions were close in magnitude ( Fig.13A). From the cluster stage to the vigorous growth stage, the growth rate of the number of leaves under each addition amount was different and decreased with the increase of the addition amount; from the vigorous growth stage to the dome stage, the growth rate of 0% addition amount was the largest (5.4 leaves) and was significantly different from other addition amounts, and the growth rates of other addition amounts were close to ( Fig.13 B). From the cluster stage to the vigorous growth stage, the growth rate of stem girth under each addition amount is different and decreases with the increase of addition amount. From the vigorous growth stage to the round-top stage, the growth rate of stem girth under each addition amount is different. The growth rate of 0% addition amount is larger (1.5cm), and the other addition amounts are relatively close ( Fig.13 C).

[0124] The plant height growth rate from the cluster stage to the vigorous growth stage of each treatment was higher than that from the vigorous growth stage to the dome stage. From the cluster stage to the vigorous growth stage, the plant height growth rate generally decreased with the increase in the amount of fermented substrate added; from the vigorous growth stage to the dome stage, the plant height growth rate also decreased with the increase in the amount added ( Fig.13 D). The leaf growth rate from the cluster stage to the vigorous growth stage of each treatment was higher than the leaf growth rate from the vigorous growth stage to the dome stage. From the cluster stage to the vigorous growth stage, the leaf growth rate generally decreased with the increase of the amount of fermented substrate added; from the vigorous growth stage to the dome stage, the leaf growth rate also decreased with the increase of the amount added ( Fig.13 E). The stem girth growth rate from the cluster stage to the vigorous growth stage is generally higher than the stem girth growth rate from the vigorous growth stage to the dome stage. From the cluster stage to the vigorous growth stage, the stem girth growth rate generally decreases with the increase in the amount of fermented substrate added from fungus and medicinal residues; from the vigorous growth stage to the dome stage, the stem girth growth rate also decreases with the increase in the amount added ( Fig.13 F).

[0125] 2.3 Effects of each treatment on economic traits of flue-cured tobacco

[0126] As the addition amount of the fermented substrate of fungus and medicinal residues increased from 0% to 15%, the various indicators of the economic traits of flue-cured tobacco showed different trends of change (Table 5). The yield showed an upward trend, which fully demonstrated that the fermented substrate of fungus and medicinal residues had a promoting effect on the yield. The overall output value showed a fluctuating upward trend. It was affected by the combined effects of multiple factors such as yield, average price and tobacco leaf grade structure. Although it had a positive impact on the output value, it was not a simple linear increase relationship. The average price showed a fluctuating change characteristic, and no significant difference was shown among the treatments. The effect of 5% on the rate of top-grade tobacco was not significant, but it decreased significantly at 10% and 15%, which may be due to the impact on the quality of tobacco leaves, resulting in a decrease. The change trend of the rate of top-medium tobacco was similar to that of top-grade tobacco. In summary, in actual production, it is necessary to comprehensively consider multiple indicators such as yield, output value, average price and tobacco rate, and reasonably select the addition amount of the fermented substrate of fungus and medicinal residues to maximize the economic benefits of flue-cured tobacco and optimize the quality of tobacco leaves. At the same time, further research on its influencing mechanism will help to better guide the practice of flue-cured tobacco production.

[0127] Table 5 Variance analysis table of economic traits

[0128]

[0129] 2.4 Effects of various treatments on the chemical composition of flue-cured tobacco

[0130] The specific influence degree and change trend of different addition amounts of fungus residue fermentation matrix on the chemical components of flue-cured tobacco (Table 6) are as follows. The nicotine content was 1.8% when the addition amount was 0%, and it increased to 1.9%, 2.1%, and 2.2% as the addition amount increased to 5%, 10%, and 15%, respectively, which may indicate that it has a promoting effect on nicotine synthesis; reducing sugar was 27.6% when the addition amount was 0%, and after the addition amount increased, it dropped to 21.1%, 21.9%, and 21.0% at 5%, 10%, and 15%, respectively, showing an inhibitory effect on its accumulation; the initial value of total nitrogen was 1.2% (0% addition amount), reaching 1.3% at 5%, rising to 1.9% at 10%, and then dropping to 1.9% at 15%, showing a fluctuation trend of first rising and then falling, which is affected by multiple factors and needs further study; the initial chlorine content was 0.7% (0% addition amount), and dropped to 0.3% at 5%. , 0.6% at 10%, and 0.8% at 15%, indicating that it has a regulating effect on the chlorine content and can be reduced within a certain range; the potassium content was 1.9% and 1.9% at 0% and 5% addition, 2.5% at 10%, and 2.6% at 15%, respectively, and the changes did not show an obvious pattern and had no significant effect on it; the nitrogen-alkali ratio was 0.69 at 0% and 5%, changed to 0.89 at 10%, and 0.84 at 15%, and the fluctuation reflected the effect of the change in the ratio of total nitrogen to nicotine on the quality and combustion characteristics; the initial value of the sugar-alkali ratio was 15.2 (0% addition), and when the addition amount increased to 5%, 10%, and 15%, it dropped to 11.2, 10.3, and 9.4 respectively, reflecting the effect of the change in the ratio of reducing sugar to nicotine content on flavor and quality. In summary, different addition amounts of fungus residue fermentation substrate have significant effects on the various chemical components of flue-cured tobacco. These changes play a key role in the quality formation of flue-cured tobacco and can provide an important basis for optimizing the amount of substrate added in flue-cured tobacco production.

[0131] Table 6 Analysis results of chemical components of tobacco leaves

[0132]

[0133] Effect on growth period: Different addition amounts of fermented substrates of fungus and medicinal residues have significant effects on the growth period of flue-cured tobacco. The clustering period, vigorous growth period, budding period, topping period and harvesting and curing period are all advanced or changed with the increase of addition amount. Among them, the clustering period, vigorous growth period and topping period are significantly advanced, while the stalk killing period is not affected by the addition amount. This shows that the fermented substrate of fungus and medicinal residues can accelerate the early growth process of flue-cured tobacco, play an important role in regulating the growth and development rhythm of flue-cured tobacco, and provide a reference for the time planning and management of flue-cured tobacco planting.

[0134] Effects on agronomic traits: With the increase in the amount of fermented substrate added from fungus and medicinal residues, the plant height, leaf number, stem girth, maximum leaf length, maximum leaf width and maximum leaf area of ​​flue-cured tobacco from the cluster stage to the vigorous growth stage and then to the dome stage showed an overall upward trend, indicating that it has a promoting effect on the growth and development of flue-cured tobacco. The growth rate and growth rate of each growth stage and index are affected by the amount added. The growth rate from the cluster stage to the vigorous growth stage is higher than that from the vigorous growth stage to the dome stage, and the growth rate decreases with the increase in the amount added. This shows that the fermented substrate of fungus and medicinal residues has different effects at different growth stages of flue-cured tobacco, which provides data support for optimizing management measures during the growth process of flue-cured tobacco, such as rationally regulating fertilization, irrigation and other measures according to the amount added at different growth stages to promote better growth and development of flue-cured tobacco.

[0135] Influence of economic traits: The increase in the amount of fermented substrate of fungus and medicinal residues increased the flue-cured tobacco yield, had a positive impact on the output value but the increase was nonlinear, and was affected by the combined effect of multiple factors. The average price fluctuated but there was no significant difference among the treatments. The effect of 5% on the top-quality tobacco rate was not significant, and the trend of the upper-middle tobacco rate was similar to that of the top-quality tobacco rate. Therefore, in flue-cured tobacco production, it is necessary to comprehensively consider indicators such as yield, output value, average price and tobacco rate to select the appropriate amount of fermented substrate of fungus and medicinal residues to maximize economic benefits and optimize tobacco leaf quality, emphasizing the importance of comprehensively considering multiple economic indicators in actual production decisions.

[0136] Effect of chemical composition: Different addition amounts of the fermentation matrix of fungus and medicinal residue significantly affect the chemical composition of flue-cured tobacco. Nicotine content increases with the addition amount, which may promote synthesis; reducing sugar content decreases, inhibiting accumulation; total nitrogen first increases and then decreases in a fluctuating manner; chlorine content can be adjusted and decreases within a certain range; potassium changes without obvious regularity; nitrogen-alkali ratio and sugar-alkali ratio fluctuate, reflecting the impact on quality and combustion characteristics. These changes indicate that the fermentation matrix of fungus and medicinal residue can adjust the chemical composition of flue-cured tobacco, providing a direction for optimizing the quality of flue-cured tobacco.

[0137] The effects of the fermentation substrate of fungus and medicinal residue on the growth period, agronomic traits, economic traits and chemical composition of flue-cured tobacco may involve multiple mechanisms. It may improve the physical and chemical properties of the soil, such as soil fertility, air permeability and water retention, thereby promoting the growth and development of flue-cured tobacco. The promotion of nicotine synthesis may be related to the regulation of tobacco alkaloid synthesis pathways by certain components in the substrate; the inhibition of reducing sugar accumulation may be due to the substrate affecting the photosynthesis or carbohydrate metabolism of flue-cured tobacco. Further research on these potential mechanisms will help to more deeply understand the working principle of the fermentation substrate of fungus and medicinal residue, and provide a theoretical basis for the precise regulation of flue-cured tobacco growth and quality.

Claims

1. A method for preparing a composite fermentation fertilizer of traditional Chinese medicine residues and fungus residues, characterized in that: The steps include: (1) Pleurotus eryngii residue, Perilla leaf Chinese medicine residue and cow dung are crushed and mixed to obtain material 1; (2) adjusting the carbon-nitrogen ratio and the water content of the material 1 and then fermenting it.

2. The preparation method according to claim 1, characterized in that: The mass ratio of the Pleurotus eryngii residue, the Perilla leaf Chinese medicine residue and the cow dung in step (1) is 8-12:0.5-1.5:0.5-1.

5.

3. The preparation method according to claim 1, characterized in that: The diameter of the material after crushing in step (1) is ≤0.5 cm.

4. The preparation method according to claim 1, characterized in that: The carbon-nitrogen ratio in step (2) is adjusted to 23-27.

5. The preparation method according to claim 1, characterized in that: The water content in step (2) is adjusted to 50-70%.

6. The preparation method according to claim 1, characterized in that: The fermentation time of step (2) is 70 to 90 days, and the pile is turned every 8 to 12 days.

7. The composite fermented fertilizer of traditional Chinese medicine residue and fungus residue obtained by the preparation method according to any one of claims 1 to 6.

8. Use of the composite fermented fertilizer of traditional Chinese medicine residue and fungus residue as claimed in claim 7 in soil improvement.

9. Use of the composite fermented fertilizer of traditional Chinese medicine residue and fungus residue as claimed in claim 7 in tobacco planting.

10. Use of the composite fermented fertilizer of traditional Chinese medicine residues and fungus residues as claimed in claim 7 in cabbage planting.

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