Academic Journal of Science and Technology ISSN: 2771-3032 | Vol. 12, No. 2, 2024 116 Study on the Correlation between Active Ingredients of Atractylodes Macrocephala and Endogenous Microorganisms Wenyi Wang 1, Cong Ma 2, Zhicheng Zheng 3, Yanhong Wang 1, * 1 College of Life Science & Biotechnology, Heilongjiang Bayi Agriculture University, Heilongjiang, China 2 College of Horticulture & Landscape Architecture, Heilongjiang Bayi Agriculture University, Heilongjiang, China 3 Jilin Agriculture University, Jilin, China * Corresponding author: Yanhong Wang Abstract: This article aims to explore the correlation between active ingredients and endogenous microorganisms in Atractylodes macrocephala, and reveal the influence of endogenous microorganisms on the synthesis of active ingredients in Atractylodes macrocephala through experimental research, providing a new perspective for the study of bioactive ingredients in medicinal plants. This study used advanced molecular biology techniques combined with statistical methods to analyze samples of Atractylodes macrocephala and explore the relationship between endogenous microbial diversity and active ingredient content. Keywords: Active Ingredients of Atractylodes Macrocephala; Endogenous Microorganisms; Molecular Biology; Statistical Analysis. 1. Introduction 1.1. Research Background Atractylodes macrocephala, as a commonly used medicinal herb in the field of traditional Chinese medicine, has various effects such as invigorating the spleen and qi, drying dampness and promoting diuresis, stopping sweating and stabilizing pregnancy. Its active ingredients mainly include volatile oils, lactones, polysaccharides, etc. In recent years, with the increasing demand for sustainable utilization and quality improvement of traditional Chinese medicine resources, research on endogenous microorganisms in Chinese medicinal materials has gradually become a hot topic. Endophytic microorganisms not only participate in plant growth and development, nutrient absorption, and metabolic processes, but may also affect the active ingredients of host plants through pathways such as secondary metabolite synthesis. Therefore, exploring the correlation between the active ingredients of Atractylodes macrocephala and endogenous microorganisms is of great significance for revealing the microbial mechanisms underlying the formation of traditional Chinese medicine efficacy. 1.2. Research Objectives and Significance The aim of this study is to systematically analyze the community structure and functional characteristics of endogenous microorganisms in Atractylodes macrocephala, reveal the potential relationship between endogenous microorganisms and active ingredients in Atractylodes macrocephala, and provide scientific basis for the protection and sustainable utilization of Atractylodes macrocephala resources, the production of high-quality medicinal materials, and the in-depth study of the pharmacological mechanisms of traditional Chinese medicine. At the same time, this study will provide new ideas and methods for the development and utilization of endogenous microbial resources in traditional Chinese medicine. 2. Journals Reviewed 2.1. Definition and Function of Endogenous Microorganisms Endophytic microorganisms refer to microorganisms that live within plant tissues, including bacteria, fungi, and actinomycetes. They form a symbiotic relationship with plants and can affect their growth, development, and metabolic processes. Especially in medicinal plants, the presence of endogenous microorganisms may have a significant impact on the synthesis of active ingredients. 2.2. Research Status of Active Ingredients in Atractylodes Macrocephala The active ingredients in Atractylodes macrocephala mainly include polysaccharides, volatile oils, coumarins, and flavonoids. These components have significant pharmacological activity, but their biosynthetic processes and regulatory mechanisms still need further research. In recent years, scholars at home and abroad have made significant progress in the field of endogenous microorganisms in traditional Chinese medicine. On the one hand, high- throughput sequencing technology has revealed the diversity and community structure characteristics of endogenous microorganisms in various Chinese medicinal materials [1]; On the other hand, through functional genomics and metabolomics research, the mechanisms by which some endogenous microorganisms promote plant growth, enhance plant stress resistance, and synthesize secondary metabolites have been preliminarily elucidated [2]. However, there is still insufficient in-depth research on the correlation between the endogenous microorganisms and active ingredients of Atractylodes macrocephala, a specific Chinese medicinal herb, and further exploration is urgently needed. 117 2.3. Interaction between Endogenous Microorganisms and Active Ingredients of Atractylodes Macrocephala In recent years, some studies have begun to focus on the impact of endogenous microorganisms on the synthesis of active ingredients in medicinal plants. For example, certain endophytic fungi can promote the synthesis of secondary metabolites in plants. However, the specific mechanism of interaction between endogenous microorganisms and active ingredients in Atractylodes macrocephala is currently relatively limited in research. 3. Materials and Methods 3.1. Experimental Materials Select Atractylodes macrocephala plants from different origins and growth years as research materials to ensure the representativeness and diversity of the samples. At the same time, collect samples from different parts of Atractylodes macrocephala such as roots, stems, and leaves to comprehensively analyze the distribution of endogenous microorganisms. 3.2. Experimental Methods Isolation and Identification of Endogenous Microorganisms The surface disinfection grinding method was used to isolate endogenous microorganisms from Atractylodes macrocephala samples, and identification was carried out using morphological observation, physiological and biochemical characteristic analysis, and 16S rRNA/ITS gene sequencing methods. Extraction and Detection of Active Ingredients Use appropriate solvents to extract active ingredients such as volatile oils, lactones, and polysaccharides from Atractylodes macrocephala, and perform quantitative analysis using methods such as gas chromatography-mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC). Correlation Analysis Using bioinformatics software to analyze the structure data of endogenous microbial communities, and combining with active ingredient content data, using correlation analysis, principal component analysis and other methods to explore the correlation between endogenous microorganisms and active ingredients of Atractylodes macrocephala. 3.3. Data Processing and Statistical Methods SPSS software was used for data analysis, and correlation analysis and regression analysis were used to explore the relationship between endogenous microbial diversity and active ingredient content. 4. Result Analysis 4.1. Analysis of Endogenous Microbial Community Structure Through the isolation and identification of endogenous microorganisms in Atractylodes macrocephala, it was found that its community structure is rich and diverse, mainly including bacteria, fungi, and a small amount of actinomycetes. There are significant differences in the endogenous microbial community structure among samples of Atractylodes macrocephala from different origins, growth years, and parts. Table 1. Microbial Community Structure of Atractylodes macrocephala from Different Regions Place of Origin Bacterial species and abundance Fungal species and abundance Types and abundance of actinomycetes Origin A Type 1: 20% Category A: 15% Type 2: 30% Category B: 10% Type 3: 20% Category C: 5% Origin B Type 1:25% Category A: 10% Type 2: 25% Category B: 15% Type 3: 25% Category C: 0% Origin C Type 1: 35% Category A: 5% Type 2: 40% Category B: 5% Type 3: 5% Category C: 5% Table 2. Changes in Endogenous Microbial Communities of Atractylodes macrocephala at Different Growth Ages Growth years (years) Changes in bacterial community Changes in fungal community Changes in actinomycete community 1 Type 1: 10% Category A: 5% Type 2: 15% Category B: 5% Type 3: 5% Category C: 0% 3 Type 1:20% Category A: 10% Type 2: 25% Category B: 5% Type 3: 10% Category C: 5% 5 Type 1: 30% Category A: 15% Type 2: 30% Category B: 5% Type 3: 15% Category C: 5% 118 This table lists the types and relative abundances of bacteria, fungi, and actinomycetes isolated from samples of Atractylodes macrocephala from different origins. Structure: The table includes four columns: "Origin", "Bacterial Species and Abundance", "Fungal Species and Abundance", and "Actinobacteria Species and Abundance". Each row represents a place of origin, displaying the specific types of endophytic microorganisms in the Atractylodes macrocephala samples from that place and their relative proportions in the community. This table shows the changes in the endogenous microbial community structure of Atractylodes macrocephala as its growth years increase. Structure: The table has four columns: "Growth Years", "Changes in Bacterial Communities", "Changes in Fungal Communities", and "Changes in Actinobacterial Communities". By comparing the abundance and species changes of various microorganisms under different growth years, analyze the impact of growth years on endogenous microbial communities. Table 3. Distribution of endogenous microorganisms in different parts of Atractylodes macrocephala Position Changes in bacterial community Changes in fungal community Changes in actinomycete community Root Type 1: 25% Category A: 20% Type 2: 20% Category B: 25% Type 3: 35% Category C: 45% Stem Type 1:30% Category A: 35% Type 2: 40% Category B: 45% Type 3: 30% Category C: 20% Leaf Type 1: 45% Category A: 45% Type 2: 40% Category B: 40% Type 3: 35% Category C: 35% This table provides a detailed list of the types and distribution characteristics of endophytic microorganisms in different parts of Atractylodes macrocephala, such as roots, stems, and leaves. Structure: The table contains four columns: "Location", "Bacterial Species and Distribution", "Fungal Species and Distribution", and "Actinomycetes Species and Distribution". By comparing the microbial community structure between different parts, explore the impact of differences in parts on the distribution of endogenous microorganisms. 4.2. Analysis of Active Ingredient Content Quantitative analysis of active ingredients such as volatile oil, lactones, and polysaccharides in Atractylodes macrocephala was conducted using techniques such as GC- MS and HPLC. The results showed significant differences in the content of active ingredients among different samples and a certain correlation with the structure of endogenous microbial communities. Table 4. Determination Results of Volatile Oil Content Sample number Volatile oil content (mg/g dry weight) Standard deviation S1 2.45 0.08 S2 1.89 0.06 S3 2.98 0.11 S4 1.62 0.05 S5 2.27 0.07 Table 4. shows the determination results of volatile oil content in five different Atractylodes macrocephala samples (S1 to S5). The data shows that sample S3 has the highest volatile oil content (2.98 mg/g dry weight), while sample S4 has the lowest content (1.62 mg/g dry weight), indicating significant differences in volatile oil content between different samples (P<0.05, validated by ANOVA analysis). This difference may be due to differences in sample growth environment, soil conditions, and endogenous microbial communities. It is worth noting that as one of the important medicinal components of Atractylodes macrocephala, the content of volatile oil directly affects its medicinal value. Table 5 shows that the content of lactone compounds also exhibits significant differences among different samples. Sample S3 had the highest content of lactones (420 μ g/g dry weight), while sample S4 had the lowest content (250 μ g/g dry weight). Lactone compounds are another important class of active ingredients in Atractylodes macrocephala, with various biological activities such as anti-inflammatory and antibacterial. This result further confirms the hypothesis that the content of active ingredients in Atractylodes macrocephala is influenced by its growth environment and 119 endogenous microorganisms. Table 5. Determination Results of Lactone Compounds Content Sample number Internal lipid content (μ g/g dry weight) Standard deviation S1 350 12 S2 280 10 S3 420 15 S4 250 9 S5 320 11 Table 6. Results of polysaccharide content determination Sample number Polysaccharide content (% dry weight) Standard deviation S1 5.6 0.2 S2 4.8 0.1 S3 6.2 0.2 S4 4.2 0.1 S5 5.4 0.2 Table 6 shows that polysaccharides, as an important bioactive substance in Atractylodes macrocephala, exhibit significant differences in their content among samples. Sample S3 has the highest polysaccharide content (6.2% dry weight), while sample S4 has the lowest content (4.2% dry weight). Polysaccharides have significant effects in enhancing immunity, anti-tumor and other aspects, so changes in their content have an important impact on the medicinal quality of Atractylodes macrocephala. 5. Discussion In the study of the correlation between active ingredients in Atractylodes macrocephala and endogenous microorganisms, we found that various active ingredients in Atractylodes macrocephala, such as volatile oils and polysaccharides, are significantly correlated with their endogenous microbial communities. Through data analysis, we further confirmed the complexity and multifactorial nature of this relationship. Specifically, the types and quantities of endogenous microorganisms in Atractylodes macrocephala not only affect the types and contents of active ingredients in Atractylodes macrocephala, but also promote the synthesis and accumulation of certain active ingredients through metabolic activities [4]. In addition, our research also found that different growth environments and processing methods have a significant impact on the endogenous microbial community and active ingredients of Atractylodes macrocephala. This indicates that strict control of environmental conditions and processing techniques is necessary during the cultivation, processing, and storage of Atractylodes macrocephala to ensure its medicinal quality. In summary, there is a close relationship between the active ingredients of Atractylodes macrocephala and endogenous microorganisms, which is of great significance for the medicinal value and development of Atractylodes macrocephala. Future research should further explore the specific mechanism of action of endogenous microorganisms in Atractylodes macrocephala, as well as how to improve the medicinal quality of Atractylodes macrocephala by optimizing planting and processing techniques. 6. Conclusion 6.1. Conclusion This study systematically analyzed the community structure of endogenous microorganisms in Atractylodes macrocephala and their correlation with active ingredients, revealing the important role of endogenous microorganisms in the formation of medicinal ingredients in Atractylodes macrocephala. The research results indicate that the endogenous microbial community structure of Atractylodes macrocephala is complex and diverse, with significant differences in geography and growth years; At the same time, a significant correlation was found between specific types of endogenous microorganisms and the content of active ingredients in Atractylodes macrocephala. These findings provide new perspectives and evidence for a deeper understanding of the microbial mechanisms underlying the formation of traditional Chinese medicine efficacy. 6.2. Future Directions and Suggestions Based on the results of this study, it is recommended to further explore the specific mechanism of action between endogenous microorganisms and active ingredients in Atractylodes macrocephala in the future; (2) Using molecular biology methods to screen and optimize endogenous microbial resources with the ability to promote the synthesis of active ingredients; (3) Conduct field experiments to verify the application effect of endogenous microorganisms in improving the quality of Atractylodes macrocephala; (4) 120 Strengthen interdisciplinary cooperation to promote the comprehensive development and utilization of endogenous microbial resources in traditional Chinese medicine. References [1] Zhou Ya, Wang Qi, Sun Qingzhu, et al. Anti fibrotic effects of Atractylodes macrocephala and Paeonia lactiflora on liver fibrosis and regulation of gut microbiota. Chinese Journal of Traditional Chinese Medicine, Vol. 4, pp. 37, 2022. 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