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Article

A Chinese Herbal Compound Fertilizer Improved the Soil Bacterial Community and Promoted the Quality of Chrysanthemum morifolium ‘Huangju’

by
Hongliang Li
1,
Hongyao Qu
1,
Huaqiang Xuan
2,
Bei Liu
3,4,
Lixiang Zhu
1,
Xianchao Shang
1,
Yi Xie
1,
Li Zhang
1,
Long Yang
1,
Ling Yuan
5,
Sitakanta Pattanaik
5,
Li Xiang
6 and
Xin Hou
1,*
1
College of Plant Protection, Shandong Agricultural University, Tai’an 271000, China
2
Liaocheng Academy of Agricultural Sciences, Liaocheng 252000, China
3
Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China
4
Wuhan Kindstar Zhenyuan Medical Laboratory Co., Ltd., Wuhan 420076, China
5
Department of Plant and Soil Sciences, Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, KY 40546, USA
6
Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
*
Author to whom correspondence should be addressed.
Agronomy 2025, 15(7), 1512; https://doi.org/10.3390/agronomy15071512
Submission received: 26 March 2025 / Revised: 5 June 2025 / Accepted: 17 June 2025 / Published: 21 June 2025
(This article belongs to the Section Innovative Cropping Systems)
Browse Figures
Versions Notes

Abstract

Chrysanthemum morifolium, ‘Huangju’, is a golden chrysanthemum used for making tea. Limited by land resources, the continuous cropping of Chrysanthemum morifolium ‘Huangju’ has led to serious soil issues, which affects its yield and quality. In this study, different ratios of traditional Chinese medicine compound fertilizers were used to regulate the soil environment in order to achieve the green prevention and control of continuous cropping obstacles of the golden chrysanthemum. Five treatments were set up in the experiment: the control (CK) and different proportions of the Chinese herbal compound fertilizer T1, T2, T3, and T4. After the application of the traditional Chinese medicine compound fertilizer, the physical and chemical soil properties of the golden chrysanthemum were changed to varying degrees, resulting in an increased yield of golden silk chrysanthemum and an improved tea quality. This preliminary study on the application of the traditional Chinese medicine compound fertilizer T2 and T3—that is, Sophora flavescens–Stemona sessilifolia–Mentha haplocalyx–Perilla frutescens–Artemisia annua at ratios of 2:1:2:1:1.5 and 3:1:3:1:2—treatments provided the best results and can be further developed to alleviate the continuous cropping obstacles of fertilizers.

1. Introduction

Chrysanthemum morifolium, ‘Huangju‘ (golden Chrysanthemum), contains a variety of flavonoids and other components, with the effect of calming the liver and improving eyesight, clearing away heat, and detoxifying. It is a popular chrysanthemum for tea, and the market demand is increasing year by year [1]. However, due to the limitation of land resources, the continuous cropping of golden chrysanthemum is very common and has led to continuous cropping problems, which seriously affect its yield and quality [2].
The traditional control method for the continuous cropping obstacle is to use chemical agents for soil fumigation or spraying. Chemical fumigants have a broad-spectrum bactericidal effect, which not only inhibits fungi but also inhibits bacteria and actinomycetes, changing their diversity and dominant populations [3].
Based on the analysis of the components of chemical fumigants and the components contained in a variety of plants and their decomposition products, researchers found that many plant decomposition products can also achieve the fumigation and sterilization of chemical fumigants in soil, resulting in the soil treatment method of biological fumigation. Biofumigation is a method to inhibit or kill pests in soil by using volatile gases or bactericidal compounds produced during the decomposition of plant-derived organic matter. Biological fumigation can effectively control soil-borne diseases, improve the soil environment, and increase the soil organic matter content [4].
Compared with ordinary plants, botanical Chinese medicinal materials contain more types and higher contents of bioactive components. Bioactive substances and secondary metabolites contained in Chinese herbal medicines and their decomposition products can be used to manipulate the bacteria and fungi in the soil and also have a stronger biological fumigation effect than ordinary plants. The alkaloids of Sophora flavescens and Stemona sessilifolia can effectively inhibit or kill pests in the field. The extracts of Mentha haplocalyx Briq. and Perilla frutescens have obvious antibacterial effects. Artemisia annua inhibits the seed germination and seedling growth of weeds in the field [5].
In this study, through field experiments and pot experiments, traditional Chinese medicine compound fertilizers prepared with different proportions of Sophora flavescens–Stemona sessilifolia–Mentha haplocalyx–Perilla frutescens–Artemisia annua were used to determine their impact on soil nutrients, the soil microbial quantity and species, and the growth and development of Chrysanthemum morifolium, ‘Huangju’. We studied the physiological mechanism of applying traditional Chinese medicine compound fertilizers as biological fumigation materials to regulate the biological characteristics of the soil of Chrysanthemum morifolium, ‘Huangju’, and promote the growth of Chrysanthemum morifolium ‘Huangju’.

2. Materials and Methods

2.1. Overview of Test Materials

The seedling cuttings of golden silk chrysanthemum were obtained from the self-cultivated seedling cuttings in the experimental base of Shandong Agricultural University, Tai‘an City, Shandong Province, China; the Chinese herbal medicines Sophora flavescens–Stemona sessilifolia–Mentha haplocalyx–Perilla frutescens–Artemisia annua, used in Chinese herbal compound fertilizer materials, were purchased from Jiaoyu Traditional Chinese Medicine Plantation Base, Daiyue District, Tai’an City, Shandong Province, China.
The field experiment was carried out in Jiaoyu Chinese herbal medicine planting base in Daiyue District, Tai‘an City, Shandong Province, China. The soil properties tested in the field experiment are as follows: organic matter (16.67 g kg−1), total nitrogen (3.17 g kg−1), total phosphorus (56.31 g kg−1), total potassium (88.32 mg kg−1), available nitrogen (29.43 mg kg−1), available phosphorus (9.22 mg kg−1), and available potassium (15.06 mg kg−1).

2.2. Experimental Design

The field experiment was conducted in a randomized block design with a plot area of 1 × 10 m2 and 3 replicates. The base fertilizer is compound fertilizer (16-18-6) 337.5 kg hm−2, the ratio of traditional Chinese medicine compound fertilizer is shown in Table 1, and the dosage is 500 kg per hectare. The base fertilizer and traditional Chinese medicine compound fertilizer were evenly spread on the surface and then turned into the soil. On 16 April 2021, the plant height of 15 ± 2 cm was selected and transplanted according to a row spacing of 30 cm and a plant spacing of 25 cm. All treatments adopted the same management measures during the growth period.

2.3. Sampling Method

Soil sampling: Soil samples were collected via five-point sampling method on November 10 (full-bloom stage). Some of them were taken back to the laboratory for natural drying and grinding for the determination of soil organic matter, available nitrogen, phosphorus and potassium. At the same time, chrysanthemum root soil was collected via shaking root method and stored at −80 °C for microbial community structure analysis. The determination of soil physical and chemical properties was conducted with three replicates for each treatment.
Sampling of aboveground dry matter accumulation and nutrient absorption, yield and quality determination: On 1 November 2021, 2 m2 of golden silk chrysanthemum was randomly selected from each plot for marking. On 10 November 2021, golden silk chrysanthemum was picked, and the open inflorescence was picked to determine the inflorescence properties.

2.4. Determination Items and Methods

2.4.1. Physiochemical Properties of Soil

Soil organic matter, available nitrogen, available phosphorus and available potassium were determined using potassium dichromate volumetric method, alkali solution diffusion method, sodium bicarbonate-molybdenum antimony colorimetric method, and flame spectrophotometer method, respectively.

2.4.2. Extraction of Soil DNA

Total microbial DNA was extracted from soil samples using a kit. DNA extraction quality was assessed by 0.8% agarose gel electrophoresis, and DNA concentration was quantified using an ultraviolet spectrophotometer. PCR amplification of the V3–V4 region of the 16S rDNA was performed using primers *341F*: 5′-CCTACGGGNGGCWGCAG-3′ and *805R*: 5′-GACTACHVGGGTATCTAATCC-3′. The amplification system was prepared using the high-fidelity enzyme Phusion High-Fidelity PCR Master Mix with HF Buffer (PCR Mix Buffer: 25 μL, DMSO: 3 μL, F/R primer pair: 3 μL, gDNA: 10 μL, Nuclease-free water: 50 μL). The TruSeq Nano DNA LT Library Prep Kit (Illumina, San Diego, CA, USA) was used to prepare the sequencing library. Trimmomatic (v. 0.39) was used for raw data quality control, and FLASH (v. 1.2.11) was used to merge quality-controlled sequences. According to the closed-reference clustering method, VSEARCH (v. 2.13.7) was used to cluster the merged FASTA sequences, removing chimeras during clustering to obtain OTU representative sequences. Optimized sequences were mapped to the OTU representative sequences using a similarity threshold of >97% to generate and annotate OTUs.

2.4.3. Yield and Quality

The randomly marked plants of Chrysanthemum morifolium ‘Huangju’ were picked, the number of inflorescences per plant was recorded, the inflorescence diameter was measured, the fresh weight of a single inflorescence was weighed, and the yield of Chrysanthemum morifolium ‘Huangju’ was calculated. Fresh flower weight per plant was used as the yield standard per plant. Forty-five replicates per treatment were selected based on the experimental planting protocol.
Inflorescence soluble sugar content was determined via anthrone colorimetry. Total amino acid content was determined through ninhydrin solution coloration method. Nitrogen content was determined via Kjeldahl method. Protein content = nitrogen content × 6.25. Carotenoid content was determined using the 95% ethanol extraction method.
The flavonoid content in inflorescence was determined by sodium nitrite–aluminum nitrate colorimetry. Total phenols were determined using Folin-Ciocalteu method [6]. The contents of chlorogenic acid, luteoloside and 3,5-O-dicaffeoylquinic acid in inflorescences were determined according to the method specified in the 2020 edition of the Pharmacopoeia of China.

2.4.4. Data Processing

DPS2019 software LSD method was used for data statistical analysis, and Microsoft Excel 2013 was used to sort out the data and draw charts.

3. Results

3.1. Effects of Chinese Herbal Compound Fertilizer on Soil Nutrients

The soil-available nitrogen content of each treatment was higher than that of CK, with T1 and T3 showing the highest levels at flowering (Figure 1A). The application of the traditional Chinese medicine compound fertilizer increased soil-available phosphorus content. The soil-available phosphorus content of T3 was the highest (Figure 1B). The soil-available potassium content in T2, T3 and T4 treatments was significantly higher than in other treatments (Figure 1C). Compared to the control, the application of Chinese herbal compound fertilizer significantly increased the soil organic matter. The content of soil organic matter in T1, T2, T3 and T4 increased with the increase in the proportion of Sophora flavescens and Mentha haplocalyx in the flowering period, and the content of T4 was the highest (Figure 1D).

3.2. Effects of Chinese Herbal Compound Fertilizer on Soil Microorganisms of Chrysanthemum morifolium cv. Jinsihuangju

3.2.1. Effects on Soil Microbial Community Composition

Effects on the Relative Abundance of Bacterial Phylum Level
The Chao1 and ACE indices of bacteria in T2 and T3 increased by 1.41–1.87% and 1.09–2.89%, respectively, compared to the control, indicating that T2 and T3 improved soil bacterial abundance. Conversely, the Chao1 and ACE indices of bacteria in T1 and T4 were significantly lower than the control, reducing soil bacterial abundance. The application of traditional Chinese medicine fertilizer increased the Shannon and Simpson indices, although the differences from the control were not significant, suggesting enhanced soil bacterial diversity (Table S1). In the first principal coordinate component, the distance between CK and each treatment with Chinese medicine fertilizer was significant. In the third principal coordinate component, the distances between T1, T2 and T3 were significant, while the distance between T4 and T3 was not significant. This indicates that applying Chinese medicine fertilizer significantly altered soil bacterial richness and community composition, with different ratios significantly affecting these parameters (Figure S1).
A total of 50 bacterial phyla were detected. The bacterial phyla with a relative abundance of more than 1% include Proteobacteria, Acidobacteriota, Actinobacteriota, Gemmatimonadota, Chloroflexi, Bacteroidota, Myxococcota, Planctomycetota, Verrucomicrobiota, Patescibacteria and Methylomirabilota. Proteobacteria, Acidobacteria and Actinobacteria are the three dominant phyla. The effects of Chinese herbal compound fertilizer on soil microbial groups varied with the formulation. Chinese herbal compound fertilizer application increased the relative abundance of Proteobacteria, and the relative abundance of T3 was the highest, followed by T4. The relative abundance of Acidobacteria was the lowest in T2 and the highest in T4. The relative abundance of Actinobacteria in T2 was the highest, followed by T4, which increased by 10.69% and 0.69%, respectively, compared with CK. In addition, the relative abundance of Gemmatimonadetes in CK was 10.12%, which was higher than that of all treatments applying Chinese herbal compound fertilizer (Figure 2).
Effect on the Relative Abundance of Bacteria
Significant differences in bacterial genera were observed in treatments. The relative abundance of Vicinamibacteraceae was the highest in all Chinese herbal compound fertilizer treatments, while the relative abundance of RB41 was the highest in CK. The relative abundance of Vicinamibacteraceae in T4 was the highest, 4.66%, while in T2, it was 6.26% lower than in CK. The application of Chinese herbal compound fertilizer reduced the relative abundance of RB41; T4 had the highest relative abundance. Compared with CK, the application of traditional Chinese medicine compound fertilizer increased the relative abundance of Lysobacter and Pseudomonas. T4 had the highest relative abundance of Lysobacter. The relative abundance of T2 Pseudomonas was the highest (Figure 3).

3.3. Effect on Dry Matter Accumulation of Aboveground Part

The aboveground (shoot) dry weight from the T3 treatment was significantly higher than that of other treatments, and the T1 treatment had the lowest aboveground dry weight (Figure 4A). The underground (root) dry weight in the T3 treatment was significantly higher than that in the other treatments, with no significant difference between the other treatments (Figure 4B).

3.4. Effect on Nitrogen Accumulation

Different Chinese herbal compound fertilizer formulations affected stem nitrogen accumulation in Chrysanthemum morifolium ‘Huangju′ and increased nitrogen accumulation in leaves and inflorescences. T2 and T3 increased stem nitrogen accumulation by 19.80% and 5.86%, respectively, compared to the control, while T1 and T4 showed decreases of 3.39% and 15.39%, respectively (Figure 5A). The application of traditional Chinese medicine compound fertilizer increased Chrysanthemum morifolium ‘Huangju’ leaf nitrogen accumulation by 9.21–61.53% compared with the control, following the order T3 > T4 > T2 > T1 > CK (Figure 5B). The application of traditional Chinese medicine compound fertilizer increased the inflorescence nitrogen accumulation by 28.57–82.56% compared with the control, following the order T3 > T2 > T4 > T1 > CK (Figure 5C).

3.5. Effect on Phosphorus Accumulation

Different Chinese herbal compound fertilizer formulations affect the phosphorus accumulation in the stems of Chrysanthemum morifolium. Stem phosphorus accumulation in T1 and T4 was significantly higher than in the control, which was 24.47% and 56.74% higher than that of the control, respectively (Figure 6A). Leaf phosphorus accumulation followed the order T3 > T1 > T2 > T4 > CK (Figure 6B).
Phosphorus accumulation in the inflorescence of T1 and T2 was 38.81% and 24.26% lower than that of the control, respectively. The phosphorus accumulation in the inflorescence of T3 and T4 was 21.75% and 114.24% higher than that of the control, respectively (Figure 6C).

3.6. Effect on Potassium Accumulation

The application of traditional Chinese medicine compound fertilizer increased the potassium accumulation in stems and inflorescences, and the potassium accumulation of stems increased by 28.69–76.87% compared with the control. The potassium accumulation in the stems of T2 was the highest. The potassium accumulation in inflorescence increased by 27.71–73.82% compared with the control (Figure 7A).
The potassium accumulation in leaves of T1 and T2 was 18.65% and 15.86% lower than the control, respectively, while that of T3 and T4 was 23.14% and 4.63% higher than the control, respectively. The potassium accumulation in leaves of T3 was the highest (Figure 7B). The potassium accumulation of T3 inflorescence was the highest (Figure 7C).

3.7. Effect of Traditional Chinese Medicine Compound Fertilizer on the Yield of Golden Silk Chrysanthemum

The application of Chinese herbal compound fertilizer reduced the number of inflorescences per plant, the number of open inflorescences per plant, and the opening ratio of inflorescences in T1. In contrast, T2, T3 and T4 increased these parameters.
The number of inflorescences per plant of T1 was 34.40% lower than the control, and there was no significant difference between T3 and T4 (Figure 8A). The number of single inflorescences of T2 was the highest, increased by 19.32% compared with the control and increased by 45.03–16.86% compared to T1, T3 and T4. The number of open inflorescences per plant in T1 decreased by 39.18% compared to the control, while T2, T3 and T4 showed increases of 10.97–51.72% compared with the control (Figure 8A). The number of open inflorescences per plant of T2 was the highest, which was 13.84–59.92% higher than that of T1, T3 and T4 (Figure 8B).
The inflorescence opening ratio in T1 decreased by 3.20% compared to the control, while T2, T3 and T4 showed increases of 22.49–32.37%. The inflorescence opening ratio of T2 was the highest, which was 2.23–26.87% higher than T1, T3 and T4 (Figure 8C).
The fresh weight of the single inflorescence of Chrysanthemum morifolium ‘Huangju’ increased by 3.41–34.37% compared to the control. The fresh weight of the single inflorescence of T1 was the highest, which was 11.75–23.04% higher than that of T2, T3 and T4 (Figure 8D).
In conclusion, T2 increased the number of inflorescences and the number of open inflorescences per plant, while T3 increased the opening ratio to the highest.

3.8. Effect of Chinese Herbal Compound Fertilizer on the Quality of Golden Chrysanthemum

The soluble sugar in the inflorescences treated with traditional Chinese medicine compound fertilizer was 11.22–28.90% lower than that in CK. Among the treatments, T2 had the highest soluble sugar content and T4 had the lowest. The soluble sugar content in T1 was 9.24–19.92% higher than in T2, T3 and T4 (Figure 9A).
Contrary to the change trend of soluble sugar content, the application of Chinese herbal compound fertilizer increased the protein content and amino acid content of chrysanthemum inflorescence. Protein content increased by 28.57–83.33% compared to the control, following the order T3 > T2 > T4 > T1. The content of T3 was the highest, which was 9.74–29.87% higher than that of T1, T2 and T4 (Figure 9B). The total amino acid content of T3 inflorescence was the highest, which was 105.82% higher than that of the control. Followed by T1, it increased by 29.12% compared with the control. There was no significant difference in total amino acid content between T2 and T4 inflorescences, but it was significantly lower than that of T3 (Figure 9C).
The carotenoid content in T1, T2 and T3 was 3.70–16.67% lower than the control. The carotene content of T4 was the highest, which was 9.26% higher than the control and 11.86–25.43% higher than T1, T2 and T3 (Figure 9D).
In conclusion, the application of Chinese herbal compound fertilizer reduced the soluble sugar content of chrysanthemum inflorescence and increased the protein and total amino acid content. The nutritional quality of T3 golden silk chrysanthemum inflorescence is better.
The total phenol content of T2 and T4 increased by 2.14% and 7.25%, respectively, compared to the control, and the total phenol content of T4 was the highest (Figure 10A). The flavonoid content of T1 and T2 increased by 6.98% and 3.34%, respectively, compared to the control, and the flavonoid content of T1 was the highest. The content of flavonoids in T3 and T4 decreased by 8.03% and 2.78% compared with the control (Figure 10B).
The application of traditional Chinese medicine compound fertilizer significantly increased the content of chlorogenic acid in the inflorescence of golden silk chrysanthemum. The content of chlorogenic acid in T4 was the highest. In this study, the content of chlorogenic acid in Chrysanthemum morifolium ‘Huangju’ is higher than that in the national pharmacopoeia (Figure 10C).
The content of luteoloside in the inflorescence of T1 was the lowest, which was 3.57% lower than that of the control, but there was no significant difference with the control. The content of T4 was the highest, which was 51.35–107.41% higher than that of T1, T2 and T3. This means that the content of luteoloside should not be less than 0.080%, which meets the standard (Figure 10D).
The content of 3,5-O-dicaffeoylquinic acid in the inflorescence of T1 was the highest, which was 18.60% higher than that of the control. The content of 3,5-O-dicaffeoylquinic acid in the inflorescence of T2 and T4 increased by 6.98% and 2.32%, respectively, compared with the control, and the content of T3 was the lowest, which decreased by 26.74% compared with the control. The ‘Pharmacopoeia of the People ‘s Republic of China’ requires that the content of 3,5-O-dicaffeoylquinic acid should not be less than 0.70%. The content of 3,5-O-dicaffeoylquinic acid in T3 Chrysanthemum morifolium ‘Huangju’ inflorescence is 0.63%, which is lower than the pharmacopoeia standard, and other treatments are higher than the pharmacopoeia standard (Figure 10E).

3.9. Occurrence of Root Rot of Chrysanthemum

Compared with CK, the application of Chinese herbal compound fertilizer reduced the incidence of root rot and alleviated the occurrence of the disease. T2, T3 and T4 reduced root rot by 65–70% relative to the control (Figure 11).

4. Discussion

4.1. Chinese Herbal Compound Fertilizer Improves Soil Environment

The Chinese herbal compound fertilizer used in this study is composed of five Chinese herbal medicines, including Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua. It is rich in cellulose, nitrogen, phosphorus, and potassium. It is applied to the field of Chrysanthemum morifolium ‘Huangju’ to improve soil organic matter and effective nitrogen, phosphorus and potassium components. The application of traditional Chinese medicine compound fertilizer increased the content of soil organic matter, available nitrogen and available phosphorus, which was higher than that of the control. Soil-available potassium content in T1, T2 and T3 was higher than the control during the growth period. Biofumigation can effectively improve soil physicochemical properties [7]. Due to the difference in organic matter and nitrogen, phosphorus and potassium components in Sophora flavescens/Stemona sessilifolia/Mentha haplocalyx/Perilla frutescens/Artemisia annua, the increase in soil organic matter and available nitrogen, phosphorus and potassium content by applying traditional Chinese medicine compound fertilizer in this study was different due to different formula ratios. It is speculated that the results of this study are due to the different contents of organic matter and nitrogen, phosphorus and potassium in Sophora flavescens/Stemona sessilifolia/Mentha haplocalyx/Perilla frutescens/Artemisia annua.
The application of Chinese herbal compound fertilizer increased the relative abundance of beneficial bacteria in rhizosphere soil. Both Mentha canadensis and Perilla frutescens showed antibacterial effects [7,8]. The release of artemisinin has a selective antibacterial effect on bacteria, causing changes in bacterial activity and community structure [9]. The application of Chinese herbal compound fertilizer increased the relative abundance of Proteobacteria, Actinobacteria, Vicinamibacteraceae, Lysobacter and Pseudomonas. These bacteria have the functions of decomposing plant residues, degrading complex organic matter, promoting nitrogen transformation and dissolving insoluble phosphorus in soil [10,11,12]. Whether the soil microecology is balanced is also one of the reasons for the high or low incidence of root rot in Chrysanthemum morifolium ‘Huangju’. In vitro culture shows that Pseudomonas can effectively inhibit the mycelial growth of Setosphaeria turcica [13]. Pseudomonas can produce DPAG, which is an inductive substance for the expression signal of biological self-synthesis genes and has the effect of inhibiting plant pathogens [14]. DPAG is a key substance in suppressing root diseases [15]. Among them, T3 Proteobacteria, T2 Actinobacteria and Pseudomonas had the highest relative abundance, which were conducive to soil nutrient transformation. Proteobacteria is the most abundant phylum in soil [16], and it is easily found in nutrient-rich soils [17]. After we applied Chinese herbal medicine compound fertilizer, soil fertility was significantly improved compared with CK, which may be the reason for the increase in Proteobacteria in Chinese herbal medicine compound fertilizer. The relative abundance of Proteobacteria T3 in the soil of Chrysanthemum morifolium ‘Huangju’ field was the highest, and the bacteria of Proteobacteria had the ability to decompose plant residues and fix nitrogen [18]. The relative abundance of Actinobacteria T2 was the highest, and Actinobacteria degraded cellulose and refractory aromatic compounds. Acidobacteria bacteria contain many genes encoding cellulase, which can degrade cellulose and so on [19]. With an increase in the proportion of Sophora flavescens and Mentha canadensis, the relative abundance of Acidobacteria increased, which may be one of the reasons for the change in soil organic matter. Soil bacteria promote the decomposition of traditional Chinese medicine compound fertilizer components and provide soil nutrients for the growth of Chrysanthemum morifolium ‘Huangju’. It is speculated that the Chinese herbal compound fertilizer applied to the soil releases phenols, flavonoids and alkaloids while being decomposed by bacteria and fungi in the soil. These substances belong to allelochemicals that can change the composition of soil microbial communities.
Compared to ordinary plants, plant-derived Chinese medicinal materials contain more types and higher contents of bioactive components. Utilizing the regulatory effects of nutrients, bioactive substances, secondary metabolites and their decomposed products on soil bacteria, fungi, they exhibit stronger biofumigation effects than ordinary plants. As a type of bio-organic fertilizer, Chinese herbal medicine compound fertilizers act longer than traditional chemical fertilizers. Meanwhile, they possess “pesticidal effects” or “microbial control effects” on soil microorganisms that ordinary chemical fertilizers do not have, combining the dual roles of biofumigants and compound fertilizers, which are more environmentally friendly. However, regarding cost, compound fertilizers formulated from Chinese herbal medicines are less competitive than traditional chemical fertilizers. Based on current research progress, this experiment primarily provides the possibility of a new type of environmentally friendly compound fertilizer. In terms of cost, it may be necessary to explore more effective and low-cost Chinese herbal medicines for testing. Whether the long-term use of Chinese herbal medicine compound fertilizers has adverse environmental impacts also requires verification through multi-year fertilization experiments in the future.

4.2. Chinese Medicine Compound Fertilizer Increases the Yield of Golden Silk Chrysanthemum and Improves the Quality of Golden Silk Chrysanthemum

The accumulation of dry matter and nitrogen, phosphorus and potassium nutrients had an important effect on yield formation. Nitrogen promoted the enhancement of plant photosynthetic ability, phosphorus was closely related to flower bud formation, and potassium affected the ability of photosynthetic products of reservoir organs to transport outward [20,21,22]. The dry matter accumulation and the accumulation of nitrogen, phosphorus and potassium in leaves and inflorescences of T2 and T3 plants were increased, and the accumulation of stem phosphorus was reduced, which was conducive to the transfer of phosphorus to inflorescence and the number of flower buds.
The number of inflorescences per plant and the dry weight of a single inflorescence are the main factors affecting the yield of Chrysanthemum morifolium ‘Huangju’. The number of inflorescences, the number of open inflorescences and the fresh weight of a single inflorescence of T2 and T3 were higher than the control, and the yield was significantly higher than the control. High concentrations of menthol can inhibit the germination of plant seeds to a certain extent [23]. Stemona sessilifolia may release some chemical substances into the surrounding environment. After these allelochemicals are absorbed by Chrysanthemum morifolium ‘Huangju’, they may affect the physiological metabolism of Chrysanthemum morifolium ‘Huangju’. This may be the reason for the decrease in the number of inflorescences per chrysanthemum plant in the T1 and T4 treatment groups.
Chrysanthemum morifolium ‘Huangju’ is used as a tea chrysanthemum. Its dissolution components are not only soluble sugar, protein, amino acid carotenoids and other nutrients but also flavonoids, total phenols, chlorogenic acid, luteoloside and 3,5-O-dicaffeoylquinic acid with healthcare components.
As primary metabolites, soluble reducing sugars serve not only as energy sources and structural materials (such as participating in cell wall synthesis) but also as precursors for secondary metabolism. When plants are in the rapid growth stage (such as the juvenile period) or when carbon sources are abundant, reducing sugars are mainly used for cell division, elongation, and energy storage (such as starch synthesis), and the synthesis of secondary metabolites may be inhibited. Under stress conditions (such as drought, high salinity, and pest infestations) or in the late growth stage, plants redirect more carbon resources to the synthesis of secondary metabolites to enhance their defensive or adaptive capabilities, during which the accumulation of reducing sugars may decrease [24]. Flavonoids in plants exist in the form of glycosides, which are mostly combined with monosaccharides, disaccharides or trisaccharides [25,26]. The soluble sugar content is often positively correlated with flavonoids, phenols and chlorogenic acid. Low soluble sugar content will cause flavonoids to decrease. The content of soluble sugar in T2 and T3 is higher in each treatment of Chinese herbal compound fertilizer, which is beneficial to the accumulation of secondary metabolites. Plant protein is synthesized by phenylalanine, which is produced by photosynthate through the shikimic acid metabolic pathway. The application of Chinese herbal compound fertilizer significantly reduced the content of soluble sugar in inflorescence and increased the content of proteins and total amino acids, but there was no significant difference in the content of flavonoids and total phenols compared with the control, indicating that the application of Chinese herbal compound fertilizer increased the efficiency of nitrogen transformation and promoted protein synthesis and secondary metabolism.
In this experiment, the content of T2 and T3 carotenoids was lower than the control, but there was no significant difference. The content of carotenoids and the content of 3,5-O-dicaffeoylquinic acid showed an opposite trend, which may be due to the complementary relationship between carotenoid degradation products and 3,5-O-dicaffeoylquinic acid synthesis precursors.
The secondary metabolites of Chrysanthemum morifolium ‘Huangju’ are chlorogenic acid, luteoloside and 3,5-O-dicaffeoylquinic acid. Excessive content will produce a bitter taste in tea brewing and affect the drinking experience. The contents of chlorogenic acid and luteoloside in T2 and T3 were higher than those in the 2020 edition of the ‘Pharmacopoeia‘ standard, but the contents were significantly lower than those in T4, which improved the quality of tea and the efficacy of the active ingredients.

5. Conclusions

  • The Chinese herbal medicine used in the compound fertilizer of traditional Chinese medicine is a plant source material, which is rich in organic matter. It can improve the soil organic matter content and effective nitrogen, phosphorus and potassium content in the flowering period of golden chrysanthemum, which is conducive to the nutrient absorption of golden chrysanthemum.
  • The Chinese herbal medicines selected for Chinese herbal compound fertilizers have selective bacteriostatic effects on bacteria and change the soil microflora. T2 and T3 increased the relative abundance of beneficial bacteria such as Proteobacteria and Actinobacteria and optimized the soil microflora. The relative abundance of beneficial bacteria such as Pseudomonas and Streptomyces was increased.
  • T2 and T3 increased the number of open inflorescences, the opening ratio and the fresh weight per inflorescence, thereby increasing the yield. The contents of soluble sugar, protein, total amino acids and carotenoids in T2 and T3 inflorescences and the contents of total phenols, flavonoids, chlorogenic acid, luteoloside and 3,5-O-dicaffeoylquinic acid in medicinal components were in the middle, which could not only ensure the taste but also exert a heat-clearing and detoxifying effect.
In summary, T2 and T3 Sophora flavescens/Stemona sessilifolia/Mentha haplocalyx/Perilla frutescens/Artemisia annua ratios of 2:1:2:1:1.5 and 3:1:3:1:2 treatments were the most effective in improving the soil quality, bacterial communities, and yield characteristics of Chrysanthemum morifolium ‘Huangju’.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/agronomy15071512/s1, Figure S1: Bacterial β diversity; Table S1: Species richness and diversity index of bacterial.

Author Contributions

H.L.: Writing—original draft, visualization, formal analysis; H.Q.: writing—original draft, data curation, methodology; H.X.: data curation, formal analysis, resources; B.L.: resources, methodology, supervision; L.Z. (Lixiang Zhu): methodology, conceptualization, supervision; X.S.: methodology, software, supervision; Y.X.: methodology, supervision, validation; L.Z. (Li Zhang): methodology, conceptualization, supervision; L.Y. (Long Yang): methodology, funding acquisition, project administration; L.Y. (Ling Yuan): methodology, conceptualization, supervision S.P.: methodology, conceptualization, supervision; L.X.: methodology, conceptualization, supervision; X.H.: methodology, software, project administration, writing—review and editing, visualization. All authors have read and agreed to the published version of the manuscript.

Funding

This research was supported by the Shandong Province Modern Agricultural Technology System (SDAIT-25-02).

Data Availability Statement

The original data in this study are included in the article, and further inquiries can be directed to the corresponding author.

Conflicts of Interest

Author Bei Liu was employed by the company Wuhan Kindstar Zhenyuan Medical Laboratory Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships.

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Agronomy 15 01512 g001
Figure 1. Chemical composition of the soil during the peak flowering period. (A): Soil-available nitrogen; (B): soil-available phosphorus; (C): soil-available potassium; (D): soil organic matter. Different letters in the same column indicate significant differences at 5% level.
Figure 1. Chemical composition of the soil during the peak flowering period. (A): Soil-available nitrogen; (B): soil-available phosphorus; (C): soil-available potassium; (D): soil organic matter. Different letters in the same column indicate significant differences at 5% level.
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Figure 2. Changes of relative abundance of bacterial phylum. CK: Not processed; T1: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 1:1:1:1:1; T2: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 2:1:2:1:1.5; T3: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 3:1:3:1:2; T4: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 4:0:4:0:2.
Figure 2. Changes of relative abundance of bacterial phylum. CK: Not processed; T1: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 1:1:1:1:1; T2: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 2:1:2:1:1.5; T3: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 3:1:3:1:2; T4: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 4:0:4:0:2.
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Figure 3. Changes of relative abundance of bacterial genera. CK: Not processed; T1: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 1:1:1:1:1; T2: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 2:1:2:1:1.5; T3: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 3:1:3:1:2; T4: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 4:0:4:0:2.
Figure 3. Changes of relative abundance of bacterial genera. CK: Not processed; T1: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 1:1:1:1:1; T2: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 2:1:2:1:1.5; T3: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 3:1:3:1:2; T4: Sophora flavescens:Stemona sessilifolia:Mentha haplocalyx:Perilla frutescens:Artemisia annua = 4:0:4:0:2.
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Figure 4. Dry weight of aboveground and underground parts of the plant after harvest. (A): Shoot dry weight; (B): root dry weight. Different letters in the same column indicate significant differences at 5% level.
Figure 4. Dry weight of aboveground and underground parts of the plant after harvest. (A): Shoot dry weight; (B): root dry weight. Different letters in the same column indicate significant differences at 5% level.
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Figure 5. Nitrogen accumulation in each part of the plant after harvest. (A): Stem nitrogen accumulation; (B): leaf nitrogen accumulation; (C): nitrogen accumulation of flowers. Different letters in the same column indicate significant differences at 5% level.
Figure 5. Nitrogen accumulation in each part of the plant after harvest. (A): Stem nitrogen accumulation; (B): leaf nitrogen accumulation; (C): nitrogen accumulation of flowers. Different letters in the same column indicate significant differences at 5% level.
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Figure 6. Phosphorus accumulation in each part of the plant after harvest. (A): Phosphorus accumulation in stems; (B): leaf phosphorus accumulation; (C): phosphorus accumulation in flowers. Different letters in the same column indicate significant differences at 5% level.
Figure 6. Phosphorus accumulation in each part of the plant after harvest. (A): Phosphorus accumulation in stems; (B): leaf phosphorus accumulation; (C): phosphorus accumulation in flowers. Different letters in the same column indicate significant differences at 5% level.
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Figure 7. Potassium accumulation in each part of the plant after harvest. (A): Potassium accumulation in stems; (B): potassium accumulation in leaves; (C): potassium accumulation of flowers. Different letters in the same column indicate significant differences at 5% level.
Figure 7. Potassium accumulation in each part of the plant after harvest. (A): Potassium accumulation in stems; (B): potassium accumulation in leaves; (C): potassium accumulation of flowers. Different letters in the same column indicate significant differences at 5% level.
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Figure 8. Plant yield after harvest. (A): Flower numbers; (B): open flower numbers; (C): open flower ratio; (D) single flower fresh weight; (E): flower diameter. Different letters in the same column indicate significant differences at 5% level.
Figure 8. Plant yield after harvest. (A): Flower numbers; (B): open flower numbers; (C): open flower ratio; (D) single flower fresh weight; (E): flower diameter. Different letters in the same column indicate significant differences at 5% level.
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Figure 9. Changes in nutrient composition of plant inflorescences. (A): Soluble sugar; (B): protein; (C): total amino acid; (D): carotenoid. Different letters in the same column indicate significant differences at 5% level.
Figure 9. Changes in nutrient composition of plant inflorescences. (A): Soluble sugar; (B): protein; (C): total amino acid; (D): carotenoid. Different letters in the same column indicate significant differences at 5% level.
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Figure 10. Changes in the health quality of plant inflorescences. (A): Total phenols; (B): flavonoid; (C): chlorogenic acid; (D): chlorogenic acid; (E): 3,5-O-dicaffeoyl quinic acid. Different letters in the same column indicate significant differences at 5% level.
Figure 10. Changes in the health quality of plant inflorescences. (A): Total phenols; (B): flavonoid; (C): chlorogenic acid; (D): chlorogenic acid; (E): 3,5-O-dicaffeoyl quinic acid. Different letters in the same column indicate significant differences at 5% level.
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Figure 11. Incidence of root rot. Different letters in the same column indicate significant differences at 5% level.
Figure 11. Incidence of root rot. Different letters in the same column indicate significant differences at 5% level.
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Table 1. The ratio of compound fertilizers combined with traditional Chinese medicinal materials.
Table 1. The ratio of compound fertilizers combined with traditional Chinese medicinal materials.
TreatmentsSophora
flavescens		Stemona
sessilifolia		Mentha
haplocalyx		Perilla
frutescens		Artemisia annuaannua
CK00000
T111111
T221211.5
T331312
T440402
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Li, H.; Qu, H.; Xuan, H.; Liu, B.; Zhu, L.; Shang, X.; Xie, Y.; Zhang, L.; Yang, L.; Yuan, L.; et al. A Chinese Herbal Compound Fertilizer Improved the Soil Bacterial Community and Promoted the Quality of Chrysanthemum morifolium ‘Huangju’. Agronomy 2025, 15, 1512. https://doi.org/10.3390/agronomy15071512

AMA Style

Li H, Qu H, Xuan H, Liu B, Zhu L, Shang X, Xie Y, Zhang L, Yang L, Yuan L, et al. A Chinese Herbal Compound Fertilizer Improved the Soil Bacterial Community and Promoted the Quality of Chrysanthemum morifolium ‘Huangju’. Agronomy. 2025; 15(7):1512. https://doi.org/10.3390/agronomy15071512

Chicago/Turabian Style

Li, Hongliang, Hongyao Qu, Huaqiang Xuan, Bei Liu, Lixiang Zhu, Xianchao Shang, Yi Xie, Li Zhang, Long Yang, Ling Yuan, and et al. 2025. "A Chinese Herbal Compound Fertilizer Improved the Soil Bacterial Community and Promoted the Quality of Chrysanthemum morifolium ‘Huangju’" Agronomy 15, no. 7: 1512. https://doi.org/10.3390/agronomy15071512

APA Style

Li, H., Qu, H., Xuan, H., Liu, B., Zhu, L., Shang, X., Xie, Y., Zhang, L., Yang, L., Yuan, L., Pattanaik, S., Xiang, L., & Hou, X. (2025). A Chinese Herbal Compound Fertilizer Improved the Soil Bacterial Community and Promoted the Quality of Chrysanthemum morifolium ‘Huangju’. Agronomy, 15(7), 1512. https://doi.org/10.3390/agronomy15071512

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