Analysis of the variation characteristics of Scutellaria baicalensis rhizosphere soil microorganisms at different growth stages and their relationship with the accumulation of active components

Analysis of the variation characteristics of Scutellaria baicalensis rhizosphere soil microorganisms at different growth stages and their relationship with the accumulation of active components | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Analysis of the variation characteristics of Scutellaria baicalensis rhizosphere soil microorganisms at different growth stages and their relationship with the accumulation of active components Lixia Xu, Minna Guo, Meishan Yue, Chuanyan Guo This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7476576/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In this study, Scutellaria baicalensis Gerog plants at different growth stages and their rhizosphere soil were used as experimental materials. The microbial diversity, physicochemical properties, and enzyme activities of the rhizosphere soil were systematically determined, while the active component contents of Scutellaria baicalensis shoots were analyzed. This research aims to provide a scientific basis for revealing the soil environment regulation mechanism underlying the accumulation of active components in Scutellaria baicalensis root. It revealed the association mechanisms between rhizosphere soil micro-ecological characteristics at different growth stages and active component accumulation of Scutellaria baicalensis root. The results showed that, compared with the flowering stage, the contents of total phosphorus (TP), alkali-hydrolyzable nitrogen (AN), soil organic carbon (SOC), microbial biomass carbon (MBC), and microbial biomass nitrogen (MBN), as well as the carbon/nitrogen (C/N) ratio, in Scutellaria baicalensis rhizosphere soil gradually increased during the fruiting and wilting stages. In contrast, the soil pH showed an opposite trend. Among the other soil nutrients, the contents of TP, available phosphorus (AP), and available potassium (AK) were the highest at the fruiting stage, followed by the wilting stage, the lowest at the wilting stage. Additionally, the TN content was the lowest at the flowering stage, and it was higher at the fruiting and wilting stages. Compared with the flowering stage, the activities of urease, sucrase, and alkaline phosphatase in Scutellaria baicalensis rhizosphere soil significantly increased at the fruiting and wilting stages. There are significant differences of bacterial community in Scutellaria baicalensis rhizosphere soil at different growth stages (flowering stage, fruiting stage, and withering stage). The relative abundance of bacteria at the genus level in Scutellaria baicalensis rhizosphere soil changes with the growth stages. The dominant genera in each stage is as follows: at the flowering stage, the dominant taxa are Bacillus , Geodermatophilus , and Bryobacte ; at the fruiting stage, the dominant taxa are Streptomyces , Sphingomonas , Blastococcuss , Haliangium , and Nocardioides ; At thewithering stage, the dominant taxa are Gemmatimonas , Opitutus , and Lysobacter . The relative abundance of fungi in Scutellaria baicalensis rhizosphere soil at the genus level changes with the growth stages. The dominant genera in each stage are as follows: at the flowering stage, the genera with the highest relative abundance are Arthrographis , Penicillium and Preussia ; at the fruiting stage, the genera with the highest relative abundance are Pseudogymnoascus , Aspergillus and Pseudallescheria ; at the withering stage, the genera with the highest relative abundance are Mortierella , Alternaria , Mycochlamys and Fusarium . Redundancy analysis indicated that AN and MBN were key environmental factors driving the differences of bacterial and fungal communities at the genus level in Scutellaria baicalensis rhizosphere soil at different growth stages. Pearson correlation analysis further showed that the main active components of Scutellaria baicalensis root, including baicalin, wogonoside, baicalein, and wogonin, were significantly positively correlated with the relative abundance of 10 bacterial genera such as Gemmatimonas , Opitutus , and Lysobacter , as well as with the relative abundance of 12 fungal genera such as Mortierella , Alternaria , and Mycochlamys . These results reveal that the rhizosphere microbial community plays an important role in the accumulation of root active components at different growth stages of Scutellaria baicalensis , providing a theoretical basis for the development of precision fertilization strategies and scientific management in Scutellaria baicalensis cultivation. Scutellaria baicalensis Georg High-throughput sequencing Soil microbial diversity Soil physicochemical properties Active components of roots Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Scutellaria baicalensis Georg is an important medicinal plant resource with efficacies such as clearing heat and drying dampness, purging fire and detoxifying, and cooling blood and stabilizing pregnancy (Li et al. 2022 ). The annual demand for Scutellaria baicalensis is enormous, and after years of intensive harvesting, wild Scutellaria baicalensis resources are becoming increasingly depleted. Cultivated Scutellaria baicalensis has become the main source of Scutellaria baicalensis medicinal materials (Yuan et al. 2010 ). Germplasm sources, fertilization measures, planting years, soil microecological environment and so on are the key factors affecting the quality and yield of Scutellaria baicalensis . (Slusarczyk et al. 2024 ; Li et al. 2022 a; Sun et al. 2024 ; Dong et al. 2022). Studies have found that the combined application of chemical fertilizer and bio-organic fertilizer not only significantly increases the bacterial diversity in Scutellaria baicalensis rhizosphere soil but also significantly increases the relative abundance of Actinobacteria and Chloroflexi , which is beneficial for improving the quality and yield of Scutellaria baicalensis (Sun et al. 2024 ). Additionally, the interaction between the rhizosphere microbiome and plants during plant growth also affects plant health status and productivity levels, a mechanism that is particularly critical in the medicinal plant cultivation field (Pshenichkina 2022 ; Dong et al. 2022). The plant root system, rhizosphere microorganisms, and rhizosphere soil constitute the rhizosphere micro-ecosystem. In this micro-ecosystem, biological factors such as plant genotype, plant growth and development stages, and invading pathogenic microorganisms, as well as abiotic factors such as soil composition, soil management practices, and climate conditions, influence the composition and diversity of the rhizosphere microbial community (Liu et al. 2023 ). Conversely, rhizosphere microorganisms affect plant metabolism and the composition of plant root exudates, mainly reflected in changes in plant physiological characteristics and disease resistance function, thereby influencing plant growth and development (Bai et al. 2022 ; Xiong et al. 2021 ). Rhizosphere microorganisms also have a profound impact on soil evolution, among which beneficial microorganisms play a crucial role in the transformation process of barren soil into cultivable soil. (Zhang et al. 2024 ). Thus, the interactions among plants, microorganisms, and environmental factors form a complex symbiotic network within the rhizosphere micro-ecosystem. The dynamic changes in this symbiotic network not only affect the diversity of rhizosphere microorganisms but also alter the phenotypic characteristics of plants and the accumulation patterns of active components (Lu et al. 2018 ). Some beneficial rhizosphere microorganisms, such as bacteria Gemmatimonas spp., Streptomyces spp., Nocardioides spp., and fungi Mortierella spp., Penicillium spp., Preussia spp., can enhance plant root vitality, promote plant growth, and improve the yield and root active component content of medicinal plants by participating in soil nutrient cycling, secreting growth hormones, and antimicrobial substances (Oshiki et al. 2018; Wang et al. 2022; Youseif et al. 2023 ; Pak et al. 2020; Wani et al. 2027; Al-Hosni et al. 2018 ). Conversely, harmful rhizosphere microorganisms such as fungi Alternaria spp. and Fusarium spp. can induce plant diseases, inhibit plant growth and development, and significantly reduce crop yields by infecting plant roots, secreting toxins, or competing for nutrients (Zhao et al. 2019 ; Wani et al. 2017 ). Studying the symbiotic relationships and interactions among medicinal plants, environmental factors, and microorganisms helps regulate the composition of microbial communities around the rhizosphere of medicinal plants, achieving high quality and yield of medicinal plants (Wang et al. 2022a ; Gao et al. 2024). There are few studies on the relationship between rhizosphere microbial communities and the accumulation of root active components of Scutellaria baicalensis at different growth stages under the combined application of organic and inorganic fertilizers. Given the important ecological functions and biological value of rhizosphere microorganisms, a systematic study was conducted on the variation characteristics the rhizosphere microbial communities of Scutellaria baicalensis at different growth stages. High-performance liquid chromatography (HPLC) was used to determine the content of root active components of Scutellaria baicalensis at different growth stages, and high-throughput sequencing technology was employed to analyze the rhizosphere microbial communities of Scutellaria baicalensis . This study aims to detect the microbial diversity, soil nutrient content, enzyme activity changes, and rhizosphere active component content in the rhizosphere soil of Scutellaria baicalensis at different developmental stages, exploring the associations among root active components, rhizosphere soil microorganisms, enzyme activity, and nutrients. Furthermore, it analyzes the dynamic variation characteristics of rhizosphere microorganisms during the growth and development of Scutellaria baicalensis and the accumulation patterns of active components at different growth stages. This study will provide new pathways for optimizing cultivation management measures of Scutellaria baicalensis and enhancing the yield scale and quality of medicinal plant. Experimental design and methods Description of the experimental site The experimental field is located in Fenyang County, Shanxi Province (North Latitude: 37°93′, East Longitude: 113°58′), at the experimental field of the Institute of the Industrial Crop, Shanxi Academy of Agricultural Sciences. The region has an annual average precipitation of 460 mm, an annual average temperature of ≥ 10 ℃, and a frost-free period of 170 days. The soil in the experimental field is Yuanhuang cinnamon soil, with Malan loess as the parent material and a texture of sandy loam. Experimental design. The Scutellaria baicalensis experiment began in 2023. Chemical fertilizers were urea, triple superphosphate, and potassium sulfate, containing CO(NH 2 ) 2 113 kg hm -2 , P 2 O 5 140 kg hm -2 , and K 2 O 15.8 60 kg hm -2 , respectively. The organic fertilizer was pig manure, containing H 2 O, organic mattter, total nitrogen (TN), P 2 O 5 , and K 2 O at 55%, 34.5 g kg -1 , 16.5 g kg -1 , 23.6 g kg -1 , and 15.8 g kg -1 , respectively. The fertilization treatment involved a combination of chemical fertilizers and organic fertilizer. Scutellaria baicalensis seeds were randomly sown in experimental plots of 60 square meters, with each plot containing 1300 plants. The row spacing was 30 cm, and the plant spacing was 15 cm. Soil sampling At the flowering stage, fruiting stage, and wilting stage in 2024, fifteen Scutellaria baicalensis plants were randomly selected from each replicate plot. The Scutellaria baicalensis roots with soil were shaken to remove loose soil, and the soil adhering to the root surface was brushed off with a soft brush, which was considered rhizosphere soil. The rhizosphere soil from fifteen Scutellaria baicalensis roots was mixed to form one soil sample for each treatment. Each soil sample is divided into three portions, which are used for chemical composition analysis, enzyme activity determination, and microbial diversity analysis respectively. Measurement of Scutellaria baicalensis growth indicators After collecting rhizosphere soil samples at the fruiting stage, the Scutellaria baicalensis plants were used to determine shoot fresh weight (SFW), root fresh weight (RFW), shoot dry weight (SDW) and root dry weight (RDW). Additionally, dry/fresh ratio (D/F) and root/shoot ratio (R/S), SDW/SFW and RDW/RFW were calculated. R/S = RDW/SDW. Detection of active components in Scutellaria baicalensis root High-performance liquid chromatography (HPLC) was used to determine baicalin, wogonoside, baicalein, and wogonin. Chromatographic column: kromasil eternity-5-C18 (250 mm×4.6 mm); mobile phase: acetonitrile solution (A)-0.4% acetic acid solution (B), gradient elution; column temperature: 30°C; detection wavelength: 280 nm; flow rate: 1.0 mL/min. 1.6 Measurement of soil nutrients, microbial biomass carbon, nitrogen ratio, and enzyme activity The contents of soil organic carbon (SOC), total nitrogen (TN), alkali-hydrolyzable nitrogen (AN), total phosphorus (TP), available phosphorus (AP), total potassium (TK), and available potassium (AK) were determined using the method of Bao (Bao 2000 ). The contents of soil microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN) were also measured. The activities of urease (UR), alkaline phosphatase (PP), and sucrase (SUC) were determined using the method of Guan (Guan 1986 ). Soil DNA extraction, Illumina sequencing, and data processing Using the Soil DNA Extraction Kit from Omega Bio-Tec (Norcross, GA, US), microbial genomic DNA was extracted from soil samples. The bacterial 16S rDNA V3-V4 region was amplified using BacF:ACTCCTACGGGAGGCAGCA and Bac R:GGACTACHVGGGTWTCTAAT; the fungal ITS region was amplified using ITSF (5′ GGAAGTAAAAGTCGTAACAAGG-3′)/ITSR (5′-GCTGCG TTCTTCATCGATGC-3′). PCR conditions were 98°C for 5 min, 98°C for 30 s, 53°C for 30 s, 25 cycles, 72°C for 45 s, 72°C for 5 min, and 12°C. The amplification results were subjected to 2% agarose gel electrophoresis, and the target fragments were excised and recovered using the Axygen Gel Extraction Kit. The DADA2 Method (Benjamin et al., 2016) was used to process the raw sequencing data, including primer removal, quality filtering to exclude low-quality sequences, denoising to correct noisy reads, automatic sequence merging for paired-end data, chimera detection to remove chimeric sequences, and deduplication. The deduplicated sequences generated after processing are called Amplicon Sequence Variants. Meanwhile, a feature table recording the abundance of these sequences in each sample is generated. These are the core data for subsequent microbiome analysis. Sequencing data accession numbers for bacteria and fungi in NCBI are PRJNA1303951 and PRJNA1303954, respectively. Data analysis Qime software was used to calculate Chao, Ace, and Shannon diversity index. R software was used to generate Venn Diagrams. Canoco 5.0 was used for Redundancy Analysis (RDA). Prism software was used to create bar charts. SPSS 16.0 software was used for One-Way ANOVA to analyze differences. Each experiment was repeated three times, and the measured values were expressed as Mean ± Standard Error (SE). Different letters show significant difference at P < 0.05 using Waller Duncan test. Results Changes in soil physicochemical properties of Scutellaria baicalensis at three different growth stages Changes in the physicochemical properties of Scutellaria baicalensis rhizosphere soil at three different growth stages are shown in Table 1 . Compared with the flowering stage, the contents of TK, AN, SOC,MBC, MBN, and the C/N ratio increased significantly by 6.53%, 23.86%, 31.93%, 49.01%, 31.01%, 19.52% at the fruiting stage and by 17.34%, 52.32%, 42.32%, 62.20%, 73.39%, 29.94% at the withering stage, with the order being withering stage > fruiting stage > flowering stage for all of them. The contents of TP and AP significantly increased by 38.83%, 50.68% and 27.18%, 36.59% at the fruiting stage and wilting stage compared with the flowering stage, respectively, with the order being fruiting stage > wilting stage > flowering stage. TP content was higher at the fruiting stage and wilting stage compared with the flowering stage, with little difference between the fruiting stage and wilting stage. Compared with the flowering stage, the gradual increase in contents of TN, TP, TK, AN, AP, AK, SOC, MBC, MBN, and the C/N ratio at the fruiting stage and wilting stage may be related to the gradual release of fertilizer effects after the application of organic fertilizer. pH value was the highest at the flowering stage (7.8), followed by the fruiting stage (7.5), and the lowest at the wilting stage (7.2). The gradual decrease in pH value may be related to the accumulation of rhizosphere exudates from Scutellaria baicalensis . Changes in plant mass accumulation of Scutellaria baicalensis at different growth stages Table 1 Soil physicochemical properties of Scutellaria baicalensis at different growth stages Sample pH TN g kg − 1 TP g kg − 1 (g/kg) TK g kg − 1 (g/kg) AN mg kg − 1 AP mg kg − 1 Flowering stage 7.8 ± 0.1c 1.03 ± 0.02a 1.03 ± 0.02a 13.61 ± 0.03a 43.54 ± 0.59a 33.29 ± 0.16a Fruiting stage 7.5 ± 0.1b 1.14 ± 0.01b 1.43 ± 0.03c 14.50 ± 0.02b 53.93 ± 0.99b 50.16 ± 0.12c Wilting stage 7.2 ± 0.1a 1.13 ± 0.02b 1.31 ± 0.02b 15.97 ± 0.05c 66.32 ± 0.60c 45.47 ± 0.56b Sample AK mg kg − 1 SOC g kg − 1 MBC mg kg − 1 MBN mg kg − 1 C/N -- Flowering stage 138.59 ± 1.04b 13.28 ± 0.03a 159.00 ± 0.79a 41.60 ± 0.21a 12.86 ± 0.23a -- Fruiting stage 151.87 ± 1.10c 17.52 ± 0.01b 236.93 ± 1.11b 54.50 ± 1.28b 15.37 ± 0.14b -- Wilting stage 134.28 ± 0.54a 18.90 ± 0.03c 257.90 ± 1.03c 72.13 ± 0.22c 16.71 ± 0.29c -- Values are given as the means ± SD (n = 3); Values within a row followed by different lowercase letters are significantly different (n = 3, LSD, P < 0.05) The results are shown in Fig. 1 . Compared with the flowering stage, RFW and RDW increased significantly by 60.00% and 50.99% respectively at the fruiting stage, and by 97.43% and 82.12% respectively at the withering stage. Compared with the flowering stage, SFW and RFW increased by 12.48% and 20.70% at the fruiting stage, and decreased by 41.767% and 32.26% at the wilting stage, respectively. Compared with the flowering stage, the R/S and SDW/SFW increased significantly by 25.09%, 7.31% at the fruiting stage and 168.86%, 16.32% at the withering stage, respectively; the RDW/RFW significantly decreased by 5.63% and 7.76% at the fruiting stage and wilting stage, respectively. Accumulation of active components in Scutellaria baicalensis at different growth stages As shown in Table 2 , compared with the flowering stage, the contents of four components at the fruiting stage and wilting stage increased significantly, with the following percentage increases at the fruiting stage baicalin (83.87%), wogonoside (82.00%), baicalein (48.15%), and wogonin (61.54%) and at the wilting stage baicalin (114.52%), wogonoside (84.54%), baicalein (204.94%), and wogonin (210.26%). Effects of fertilization on soil enzyme activity of Scutellaria baicalensis The enzyme activities in the rhizosphere soil at the growth and development of Scutellaria baicalensis are shown in Fig. 2 . Compared with the flowering stage, the activities of UR, PP, and SUC significantly increased by 100.67%, 29.30%, and 35.71% at the fruiting stage, and significantly increased significantly increased by 80.43%, 68.23%, and 59.07% at the wilting stage.This indicates that soil enzyme activity gradually increases with the growth stage and the gradual increase in rhizosphere soil nutrients Table 2 Contents of active components in roots at different growth stages Sample Baicalin Wogonoside Baicalein Wogonin Flowering stage 8.27 ± 0.06a 1.70 ± 0.02a 0.27 ± 0.00a 0.13 ± 0.01a Fruiting stage 15.20 ± 0.01b 3.10 ± 0.01b 0.40 ± 0.02b 0.21 ± 0.01a Wilting stage 17.73 ± 0.02c 3.14 ± 0.01c 0.82 ± 0.01c 0.40 ± 0.01c Values are given as the means ± SD (n = 3); Values within a row followed by different lowercase letters are significantly different (n = 3, LSD, P < 0.05) Effects of fertilization on soil microbial diversity of Scutellaria baicalensis We used high-throughput sequencing technology to analyze the differences in bacterial and fungal community composition in Scutellaria baicalensis rhizosphere at the flowering stage, fruiting stage, and wilting stage. Table 3 shows that the mean values of high-quality sequences of bacteria at the flowering stage, fruiting stage, and wilting stage were 53113, 55521, and 59101, respectively. Compared with the flowering stage, the high-quality sequences and Shannon index significantly increased by 4.53%, 3.06% (at the fruiting stage) and 11.27%, 4.20% (at the wilting stage), respectively. Additionally, the Chao index significantly increased by 9.85% at the wilting stage compared with the flowering stage. For fungi, the values of high-quality sequences were 46564, 43068, and 42507, respectively. Compared with the flowering stage, the Chao index and Simpson index significantly increased by 7.95% and 4.54% at the fruiting stage, and by 8.46% and 6.10% at the wilting stage. Shannon index significantly increased by 9.09% at the wilting stage compared with the flowering stage. High-quality sequences significantly decreased by 7.51% and 8.71% at the fruiting stage and wilting stage compared with the flowering stage. Table 3 Sequence numbers and diversity indices of rhizosphere soil of Scutellaria baicalensis at different growth stages Sample Chao Shannon Simpson high-quality sequences Flowering stage 3580.53 ± 80.21a 10.47 ± 0.03a 0.9982 ± 0.0001a 53113 ± 135a Fruiting stage 3601.11 ± 16.12a 10.79 ± 0.01b 0.9987 ± 0.0000a 55521 ± 89b Wilting stage 3856.21 ± 38.12c 10.91 ± 0.02c 0.9999 ± 0.0002b 59101 ± 98c Sample Chao Shannon Simpson high-quality sequences Flowering stage 288.67 ± 6.61b 5.28 ± 0.02a 0.9017 ± 0.0114a 46564 ± 70.15c Fruiting stage 265.72 ± 1.96a 5.37 ± 0.01a 0.9426 ± 0.0027b 43068 ± 48.50b Wilting stage 313.08 ± 0.93c 5.76 ± 0.06c 0.9567 ± 0.0026c 42507 ± 67.28a Values are given as the means ± SD (n = 3); Values within a row followed by different lowercase letters are significantly different (n = 3, LSD, P < 0.05) Changes in bacterial composition at the phylum level in rhizosphere soil of Scutellaria baicalensis at different growth stages In each sample, genera with relative abundance exceeding 0.5% were analyzed at the phylum level (Fig. 3 ). At the growth stage of Scutellaria baicalensis , the relative abundance of main bacterial communities in rhizosphere soil (including Actinobacteriota , Bacteroidota , Proteobacteria , and Chloroflexi significantly increased by 15.44%, 11.59%, 26.01%, and 10.88% respectively at the fruiting stage, and by 22.49%, 31.72%, 13.52%, and 22.76% respectively at the wilting stage, compared with the flowering stage. The relative abundance of Acidobacteria , Firmicutes , Verrucomicrobiot a, and Planctomycetota significantly decreased by 8.43%, 13.36%, 35.75%, and 26.82% at the fruiting stage and 16.65%, 19.82%, 57.56%, and 50.85% at the wilting stage compared with the flowering stage. The relative abundance of Gemmatimonadota significantly increased by 15.59% at the fruiting stage compared with the flowering stage, with little difference between the wilting stage and the flowering stage. At different growth stages of Scutellaria baicalensis , the relative abundance of main fungal communities in rhizosphere soil, such as Ascomycota , was 93.67%, 95.52%, and 91.41% at the flowering stage, fruiting stage, and wilting stage, respectively. The relative abundance of Mortierellomycota , Aphelidiomycota , and Chytridiomycota significantly decreased by 13.46%, 52.38%, and 25.66% at the fruiting stage and 49.20%, 63.18%, and 61.03% at the wilting stage compared with the flowering stage. The relative abundance of Basidiomycota significantly increased by 50.41% at the fruiting stage and by 207.91% at the wilting stage compared with the flowering stage. Changes in bacterial composition at the genus level in Scutellaria baicalensis rhizosphere soil at different growth stage In each sample, genera with relative abundance exceeding 0.5% were analyzed at the genus level (Fig. 4 ). Among 30 bacterial genera, Sphingomonas had the highest relative abundance, with values of 1.52%, 2.34%, and 2.07% at the flowering stage, fruiting stage, and wilting stage, respectively. Compared with the flowering stage, the relative abundance of 9 bacterial genera ( Haliangium , Blastococcus , Solirubrobacter , Streptomyces , Cellulomonas , Rubrobacter , Ensifer , Georgenia , and Iamia ) exhibited a significant increase, specifically rising by 22.74–73.58% at the fruiting stage and by 9.46–42.43% at the wilting stage. The relative abundance of these 9 genera increased from the flowering stage to the fruiting stage and decreased from the fruiting stage to the wilting stage, with the fruiting stage being higher than the flowering stage. The relative abundance of Bradyrhizobium increased by 50.23% at the fruiting stage compared with the flowering stage, with little difference between the wilting stage and the flowering stage. Compared with the flowering stage, the relative abundance of Bacillus , Nocardioides , Geodermatophilus , Mycobacterium , Bryobacter , Pseudomonas , Skermanella , Agromyces , and Devosia significantly decreased by 8.08%~31.32% at the fruiting stage and by 23.40%~57.59% at the wilting stage. Compared with the flowering stage, the relative abundance of Gemmatimonas , Lysobacter , Ramlibacter , Pedomicrobium , Rhodomicrobium , Aeromicrobium , Phenylobacterium , Opitutus , Arenimonas , and Candidatus Solibacter significantly increased by 14.76%~48.60% at the fruiting stage and by 16.34%~175.98% at the wilting stage. The relative abundance of these 10 genera gradually increased at the flowering stage, fruiting stage, and wilting stage. Among 30 fungal genera, Mycochlamy s had the highest relative abundance, with values of 4.41%, 6.78%, and 7.82% at the flowering stage, fruiting stage, and wilting stage, respectively. Compared with the flowering stage, the relative abundance of Trichocladium , Fusarium , Plectosphaerella , Cladosporium , Scopulariopsis , Sodiomyces , Mortierella , Podospora , Metarhizium , and Myrmecridium significantly increased by 24.22%~195.57% at the fruiting stage and by 43.66%~418.83% at the wilting stage.The relative abundance of these 11 genera gradually increased in rhizosphere soil during the growth stage of Scutellaria baicalensis . The relative abundance of Alternaria significantly increased by 90.67% at the wilting stage compared with the flowering stage and fruiting stage, with little difference between the flowering stage and fruiting stage. Compared with the flowering stage, the relative abundance of Preussia , Arthrographis , Eremomyces , Corynascella , Zopfiella , Bisifusarium , Penicillium , Beauveria , Solicoccozyma , and significantly decreased by 4.05%~82.83% at the fruiting stage and by 24.88%~90.58% at the wilting stage. The relative abundance of these 10 genera gradually decreased in rhizosphere soil at the growth stage of Scutellaria baicalensis . The relative abundance of Botryotrichum , Pseudogymnoascus , Aspergillus , Xenodidymella , and Subramaniula significantly increased by 28.55%~234.19% at the fruiting stage compared with the flowering stage and significantly decreased by 14.03%~47.47% at the wilting stage compared with the flowering stage. The relative abundance of these 6 genera showed a trend of first increasing and then decreasing at the three growth stages of Scutellaria baicalensis , with the relative abundance at the wilting stage being lower than that of the flowering stage. The relative abundance of Acaulium and Stolonocarpus significantly decreased by 66.07% and 75.90% at the fruiting stage compared with the flowering stage, with little difference between the flowering stage and wilting stage. Compared with the flowering stage, the relative abundance of Wardomyces and Pseudallescheria significantly increased by 43.47% and 6.53% at the fruiting stage, and by 262.99% and 64.06% at the wilting stage, respectively. Relationship between rhizosphere soil nutrients, microbial groups, and enzyme activity of Scutellaria baicalensis RDA revealed the relationship between soil chemical properties, microbial community, and enzyme activity, with soil properties as environmental factors and microbial groups as variables (Fig. 5 a, b, c, d). Axis 1 and Axis 2 explain 75.48% and 20.29% of the total variation between the community composition at the bacterial phyla level and soil properties (Fig. 5 a). The contents of AN, AP, AK, SOC, MBC, MBN, and C/N ratio were significantly positively correlated with the relative abundance of Bacteroidetes , Gemmatimonadetes , Proteobacteria , Chloroflexi , Actinobacteria , and Cyanobacteria ; The pH value was significantly positively correlated with the relative abundance of Verrucomicrobia , Firmicutes , Acidobacteria , Nitrospirae , Planctomycetes , and Armatimonadetes . The most significant factor affecting bacteria at the phyla level was AP (F = 19.8, P = 0.002), followed by MBC (F = 14.9, P = 0.014). Axis 1 and Axis 2 explain 86.70% and 4.84% of the total variation between the community composition at the bacterial genus level and soil properties (Fig. 5 b).The contents of AN, AP, AK, MBN, MBC, and C/N ratio were positively correlated with the relative abundance of Gemmatimonas , Pedomicrobium , Aeromicrobium , Arenimonas , Lysobacter , Ramlibacter , Rhodomicrobium , Phenylobacterium , Opitutus , and Candidatus Solibacter . The most significant factor affecting bacteria at the genus level was AN (F = 35.8, P = 0.002), followed by MBN (F = 30.0, P = 0.004). Axis 1 and Axis 2 explain 83.45%和15.02% of the total variation between the community composition at the fungal phyla level and soil properties (Fig. 5 c). RDA showed that the pH value and AK content were positively correlated with the relative abundance of Aphelidiomycota , Mortierellomycota , Ascomycota , and Chytridiomycota ; the relative abundance of Basidiomycota was positively correlated with the contents of AN, AP, SOC, MBC, MBN, and C/N ratio. The most significant factor affecting fungi at the phyla level was MB (F = 9.7, P = 0.018), followed by AN (F = 9.0, P = 0.018). Axis 1 and Axis 2 explain 97.28%和2.34% of the total variation between the community composition at the fungal phyla level and soil properties (Fig. 5 d).RDA showed that the contents of AN, AP, AK, MBN, MBC, and C/N ratio were positively correlated with the relative abundance of Mycochlamys , Trichocladium , Fusarium , Plectosphaerella , Cladosporium , Alternaria , Scopulariopsis , Sodiomyces , Mortierella , Podospora , Metarhizium , and Myrmecridium . The most significant factor affecting fungi at the genus level was MBN (F = 117, P = 0.004), followed by AN (F = 152, P = 0.004). Axis 1 and Axis 2 explain 95.23%和4.63% of soil enzyme activities and soil properties (Fig. 5 e). PP activity was positively correlated with the contents of AN, AP, MBC, MBN, SOC, and C/N ratio; UR activity was positively correlated with the contents of AN, AP, AK, MBC, MBN, SOC, and C/N ratio; SUC activity was positively correlated with the contents of AN, AP, MBC, MBN, SOC, and C/N ratio. The activities of the three enzymes significantly influenced soil physicochemical properties. Relationship between effective components and growth condition of Scutellaria baicalensis root and microbial community. Figures 7a and 7b are correlation heatmaps of bacterial and fungal communities with plant effective components and growth traits, respectively. The heatmaps visually present the Pearson correlation between plant indicators and microbial communities through color. Figure a shows that baicalin, wogonoside, baicalein, wogonin, SDW/SDF, and R/S were significantly positively correlated with 10 bacterial genera such as Gemmatimonas and Pedobacter , while negatively correlated with 9 bacterial genera such as Bacillus and Nocardioides ; the RDW/RFW showed the opposite trend. Figure b shows that baicalin, wogonoside, baicalein, wogonin, SDW/SFW, and R/S were significantly positively correlated with 12 fungal genera such as Mycochlamys and Trichocladium , while negatively correlated with 9 fungal genera such as Preussia and Arthrographis ; the RDW/RFW showed the opposite trend. Discussion During the cultivation of Scutellaria baicalensis , soil nutrients play an important role in promoting the formation of yield and quality. Studies have shown that nutrient cycling is closely related to soil bacterial diversity and function, especially the changes in soil nutrients at different growth stages after Scutellaria baicalensis planting, which affect the bacterial diversity in rhizosphere soil. Meanwhile, soil enzyme activity plays an important role in soil nutrient transformation. This study found that from the flowering stage to the wilting stage, urease activity in the soil gradually increased, and AN content correspondingly increased, indicating that the increase in urease activity promoted the decomposition of organic nitrogen in the soil, increasing the AN content (Table 1 ). This is consistent with the findings of Wu in Fructus aurantii (Wu et al. 2023 ). At the wilting stage, soil PP activity was higher than in the fruiting stage, while the total AP content in the soil was lower (Table 1 , Fig. 2 ). This may be because the demand for phosphorus element increases with the growth of Scutellaria baicalensis , reducing the AP content in the soil. Previous studies on Rehmannia glutinosa found that appropriate application of phosphate fertilizer increased the phosphorus content in Rehmannia glutinosa roots (Gao et al. 2021 ), supporting our result. In our previous experiments on different fertilization measures for Scutellaria baicalensis , we found that increasing organic fertilizer could improve the content of soil organic matter. Therefore, this experiment adopted the measure of combining organic fertilizer and inorganic fertilizer, which is consistent with the findings of Sun in Scutellaria baicalensis fertilization experiments (Sun et al. 2024 ). Additionally, changes in soil nutrients can alter the bacterial diversity and function in rhizosphere soil. In this study, compared with the flowering stage, the Shannon index and Chao index of the soil increased at the fruiting and wilting stages (Table 3 ), indicating that soil organic matter facilitates the formation of soil bacterial diversity. To confirm our hypothesis, we further analyzed the functions of bacteria and fungi at the phyla level during different growth stages of Scutellaria baicalensis . From the flowering stage to the withering stage, the relative abundance of Actinobacteria and Bacteroidetes gradually increased, and the contents of AN, SOC, MBC, MBN, and C/N ratio in Scutellaria baicalensis rhizosphere soil showed a gradual increasing trend (Table 1 ). The reasons are, firstly, the continuous release of fertilizer efficiency from organic fertilizer in the soil during the growth and development of Scutellaria baicalensis promotes the reproduction of nutrient-rich bacteria such as Actinobacteria and Bacteroidetes , and Actinobacteria contributes to the degradation and utilization of organic matter; secondly, from the flowering stage to the fruiting stage, the vigorous growth of Scutellaria baicalensis roots produces a large amount of rhizosphere exudates, promoting the formation of organic matter in the soil, and the rich organic matter benefits the growth of Actinobacteria . This study found that with the growth and development of Scutellaria baicalensis , the abundances of Proteobacteria and Chloroflexi in the soil both showed a trend of first increasing and then decreasing (Fig. 3 ). This variation pattern was consistent with the content dynamics of AK and AP in the soil, and also aligned with the conclusion from previous research that Proteobacteria are eutrophic bacteria (Fierer et al., 2012 ). However, this trend was not entirely consistent with the existing finding that Chloroflexi are oligotrophic bacteria (Xian et al., 2020 ). We speculate that this may vary depending on plant species. At the genus level, the study found that Bacillus spp. in the rhizosphere participates in the cycling of proteins and cellulose, and Bacillus velezensis YH-18 and Bacillus velezensis YH-20 can promote the growth of stems and leaves of peach plants and the cultivation of high-quality tree species (Shi et al. 2022 ). Geodermatophilus obcurus , isolated from dolomite marble, can tolerate harsh environments such as dryness, mitomycin C, hydrogen peroxide, ionizing radiation, and ultraviolet radiation (Montero-Calasanz et al. 2014 ). Bryobacter spp. can decompose and utilize polysaccharides, various sugars, and organic acids in carbon cycling (Li et al. 2023 ). Nocardioides spp. not only has phosphorus-solubilizing functions but also promotes the degradation of various hard-to-degrade organic compounds such as aromatic compounds, hydrocarbons, halogenated alkanes, nitrogen-containing heterocyclic compounds, and polymeric polyesters (Wang et al. 2022b ); Sphingomonas spp. can degrade 3-(4-isopropylphenyl)-1,1-dimethylurea in soils with pH above 7.0 (Bending et al. 2003 ). Streptomyces spp. participates in the decomposition of organic matter and can promote the activation of plant disease resistance mechanisms; compared with normal nitrogen fertilizer application, reducing nitrogen fertilizer by 50% combined with Streptomyces sp. NGB-Act4 and NGB-Act6 significantly promoted wheat growth and increased yield (Youseif et al. 2023 ); further demonstrating the growth-promoting effect of Streptomyces spp. on plants. Our study found that Streptomyces spp. first increased and then decreased (Fig. 4 ), indicating that this genus played an active role during the vigorous growth stage of Scutellaria baicalensis . Blastococcus spp. is widely present in soils with extreme cold, extreme heat, salinization, toxic amendments, and significant heavy metal pollution, and can be used for soil improvement (Sbissi et al. 2025 ); Haliangium can be used as a plant bactericide (Kumar et al. 2023 ). Opitutus has nitrogen fixation and nitrogen preservation functions (Hu et al. 2024 ). Lysobacter enzymogenes is a non-pathogenic strain that inhibits various crop fungal diseases by synthesizing antifungal factors (Zhao et al. 2019 ). Gemmatimonas spp. participates in the reduction of N₂O, reducing nitrogen loss in the form of N₂O, allowing more nitrogen to remain in the soil for crop absorption, thereby improving nitrogen fertilizer utilization efficiency and reducing agricultural production costs (Oshiki et al. 2022 ). The study of bacterial functions shows that the above 11 genera are related to nutrient cycling, organic matter degradation, or antagonism against soil pathogens. 4 genera had the highest abundance at the flowering stage, 4 genera had the highest relative abundance at the fruiting stage, and 3 genera had the highest relative abundance at the wilting stage. This indicates that the communities and abundance of beneficial bacteria in Scutellaria baicalensis rhizosphere at the three different growth stages show regular changes. Correlation analysis showed that Opitutus , Lysobacter , and Gemmatimonas were significantly positively correlated with the accumulation of effective components (Fig. 6 ), further supporting our results. Research on fungal functions has found that Ascomycota has higher abundance in soils with lower organic matter than in soils with higher organic matter. This phylum can participate in the formation of soil aggregates and serves as an important source of various toxins. (Li et al. 2014 ; Fu et al. 2020 ); Basidiomycota is saprophytic fungi suitable for survival in environments rich in organic matter (Fu et al. 2020 ). The results of this study showed that compared with the flowering stage, the relative abundance of Ascomycota increased at the fruiting stage and decreased at the wilting stage, while the relative abundance of Basidiomycot a gradually increased at the fruiting and wilting stages; this is related to the gradual increase in soil organic matter with the growth and development of Scutellaria baicalensis . Arthrographis spp. has the ability to produce cellulase and is commonly found in compost (Eida et al. 2011 ; Okeke et al. 1993). Some species of Penicillium spp. can produce soluble phosphorus, iron carriers, and plant hormones (such as indole-3-acetic acid and gibberellin), which play an important role in plant health (Park et al. 2020 ). Preussia spp. can produce plant growth hormones to promote plant growth (Al-Hosni et al. 2018 ). Aspergillus brunneoviolaceus HZ23 is a plant growth-promoting bacterium that changes the physicochemical properties of rhizosphere soil of pak choi and can be used as an amendment for newly reclaimed soil (Li et al. 2023 ). Pseudogymnoascus spp. is the most abundant fungal genus in the rhizosphere, and its members are usually involved in cellulose degradation (Sigler et al. 2000 ). Pseudallescheria spp. is a potential pathogen found in soil and on humans and animals (Rainer et al. 2020). Mycochlamys can participate in chitin degradation (Debode et al. 2016 ). A large amount of Mortierella alpina in the rhizosphere not only participates in the decomposition of plant residues and organic matter but also promotes plant growth by facilitating the biosynthesis of carotenoid derivatives and enhancing stress tolerance (Wani et al., 2017 ); some genera of Mortierella spp. can convert insoluble phosphorus in the soil into soluble phosphorus, improving plant phosphorus absorption capacity and promoting plant growth and development (Sang et al. 2022 ). Alternaria spp. is a potential plant pathogenic fungus that can cause potato early blight, potato brown spot, and soybean black spot (Xu et al. 2022 ). Fusarium spp. can cause crop diseases such as potato dry rot and potato Fusarium wilt (Zhang et al. 2017 ). The study of fungal functions shows that the above 10 genera are related to nutrient cycling, organic matter degradation, and plant pathogenicity. Two genera had the highest abundance at the flowering stage, 2 genera had the highest relative abundance at the fruiting stage, and 4 genera had the highest relative abundance at the wilting stage. The highest relative abundance of Mycochlamy s, Mortierella , Alternaria , and Fusarium at the wilting stage was significantly positively correlated with effective components such as baicalin (Fig. 6 ), further supporting our results. Conclusion The research results show that influenced by the growth stage, the soil physicochemical properties of Scutellaria baicalensis at different growth stages exhibit various changes. The diversity of fungi and bacteria is the highest at the wilting stage compared with fruiting stage and wilting stage. The groups of functional microorganisms among fungi and bacteria exhibit distinct characteristics at different growth stages of Scutellaria baicalensis . The association patterns between bacterial and fungal diversity and the accumulation of active components in Scutellaria baicalensis root provide key data for in-depth analysis of the ecological interaction network between plants and microorganisms. This helps explore the bidirectional mechanisms of plant metabolites regulating microbial communities and microorganisms influencing plant growth and development. It also provides a theoretical basis for optimizing plant growth and enhancing the accumulation of secondary metabolites through microbial community regulation. Declarations Author Contribution Lixia Xu and Minna Guo Wrote the main manuscript text and Meishan Yue and Chunyan Guo prepared figures 1-3. All authors reviewed the manuscript. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7476576","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":508396511,"identity":"d252c80c-4c68-4098-981d-0386a6b6040f","order_by":0,"name":"Lixia 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09:08:56","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3313570,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7476576/v1/91c143a4-b038-4fd3-a14e-839b71f3d4fe.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Analysis of the variation characteristics of Scutellaria baicalensis rhizosphere soil microorganisms at different growth stages and their relationship with the accumulation of active components","fulltext":[{"header":"Introduction","content":"\u003cp\u003e\u003cem\u003eScutellaria baicalensis\u003c/em\u003e Georg is an important medicinal plant resource with efficacies such as clearing heat and drying dampness, purging fire and detoxifying, and cooling blood and stabilizing pregnancy (Li et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The annual demand for \u003cem\u003eScutellaria baicalensis\u003c/em\u003e is enormous, and after years of intensive harvesting, wild \u003cem\u003eScutellaria baicalensis\u003c/em\u003e resources are becoming increasingly depleted. Cultivated \u003cem\u003eScutellaria baicalensis\u003c/em\u003e has become the main source of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e medicinal materials (Yuan et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Germplasm sources, fertilization measures, planting years, soil microecological environment and so on are the key factors affecting the quality and yield of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e. (Slusarczyk et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003ea; Sun et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Dong et al. 2022). Studies have found that the combined application of chemical fertilizer and bio-organic fertilizer not only significantly increases the bacterial diversity in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil but also significantly increases the relative abundance of \u003cem\u003eActinobacteria\u003c/em\u003e and \u003cem\u003eChloroflexi\u003c/em\u003e, which is beneficial for improving the quality and yield of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e (Sun et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Additionally, the interaction between the rhizosphere microbiome and plants during plant growth also affects plant health status and productivity levels, a mechanism that is particularly critical in the medicinal plant cultivation field (Pshenichkina \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Dong et al. 2022).\u003c/p\u003e\u003cp\u003eThe plant root system, rhizosphere microorganisms, and rhizosphere soil constitute the rhizosphere micro-ecosystem. In this micro-ecosystem, biological factors such as plant genotype, plant growth and development stages, and invading pathogenic microorganisms, as well as abiotic factors such as soil composition, soil management practices, and climate conditions, influence the composition and diversity of the rhizosphere microbial community (Liu et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Conversely, rhizosphere microorganisms affect plant metabolism and the composition of plant root exudates, mainly reflected in changes in plant physiological characteristics and disease resistance function, thereby influencing plant growth and development (Bai et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Xiong et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Rhizosphere microorganisms also have a profound impact on soil evolution, among which beneficial microorganisms play a crucial role in the transformation process of barren soil into cultivable soil. (Zhang et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Thus, the interactions among plants, microorganisms, and environmental factors form a complex symbiotic network within the rhizosphere micro-ecosystem. The dynamic changes in this symbiotic network not only affect the diversity of rhizosphere microorganisms but also alter the phenotypic characteristics of plants and the accumulation patterns of active components (Lu et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Some beneficial rhizosphere microorganisms, such as bacteria \u003cem\u003eGemmatimonas\u003c/em\u003e spp., \u003cem\u003eStreptomyces\u003c/em\u003e spp., \u003cem\u003eNocardioides\u003c/em\u003e spp., and fungi \u003cem\u003eMortierella\u003c/em\u003e spp., \u003cem\u003ePenicillium\u003c/em\u003e spp., \u003cem\u003ePreussia\u003c/em\u003e spp., can enhance plant root vitality, promote plant growth, and improve the yield and root active component content of medicinal plants by participating in soil nutrient cycling, secreting growth hormones, and antimicrobial substances (Oshiki et al. 2018; Wang et al. 2022; Youseif et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Pak et al. 2020; Wani et al. 2027; Al-Hosni et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Conversely, harmful rhizosphere microorganisms such as fungi \u003cem\u003eAlternaria\u003c/em\u003e spp. and \u003cem\u003eFusarium\u003c/em\u003e spp. can induce plant diseases, inhibit plant growth and development, and significantly reduce crop yields by infecting plant roots, secreting toxins, or competing for nutrients (Zhao et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Wani et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Studying the symbiotic relationships and interactions among medicinal plants, environmental factors, and microorganisms helps regulate the composition of microbial communities around the rhizosphere of medicinal plants, achieving high quality and yield of medicinal plants (Wang et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2022a\u003c/span\u003e; Gao et al. 2024).\u003c/p\u003e\u003cp\u003eThere are few studies on the relationship between rhizosphere microbial communities and the accumulation of root active components of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e at different growth stages under the combined application of organic and inorganic fertilizers. Given the important ecological functions and biological value of rhizosphere microorganisms, a systematic study was conducted on the variation characteristics the rhizosphere microbial communities of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e at different growth stages. High-performance liquid chromatography (HPLC) was used to determine the content of root active components of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e at different growth stages, and high-throughput sequencing technology was employed to analyze the rhizosphere microbial communities of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e. This study aims to detect the microbial diversity, soil nutrient content, enzyme activity changes, and rhizosphere active component content in the rhizosphere soil of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e at different developmental stages, exploring the associations among root active components, rhizosphere soil microorganisms, enzyme activity, and nutrients. Furthermore, it analyzes the dynamic variation characteristics of rhizosphere microorganisms during the growth and development of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e and the accumulation patterns of active components at different growth stages. This study will provide new pathways for optimizing cultivation management measures of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e and enhancing the yield scale and quality of medicinal plant.\u003c/p\u003e"},{"header":"Experimental design and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003eDescription of the experimental site\u003c/h2\u003e\n \u003cp\u003eThe experimental field is located in Fenyang County, Shanxi Province (North Latitude: 37\u0026deg;93\u0026prime;, East Longitude: 113\u0026deg;58\u0026prime;), at the experimental field of the Institute of the Industrial Crop, Shanxi Academy of Agricultural Sciences. The region has an annual average precipitation of 460 mm, an annual average temperature of \u0026ge;\u0026thinsp;10 ℃, and a frost-free period of 170 days. The soil in the experimental field is Yuanhuang cinnamon soil, with Malan loess as the parent material and a texture of sandy loam.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eExperimental design.\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe \u003cem\u003eScutellaria baicalensis\u003c/em\u003e experiment began in 2023. Chemical fertilizers were urea, triple superphosphate, and potassium sulfate, containing CO(NH\u003csub\u003e2\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003e 113 kg hm\u003csup\u003e-2\u003c/sup\u003e, P\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e 140 kg hm\u003csup\u003e-2\u003c/sup\u003e, and K\u003csub\u003e2\u003c/sub\u003eO 15.8 60 kg hm\u003csup\u003e-2\u003c/sup\u003e, respectively. The organic fertilizer was pig manure, containing H\u003csub\u003e2\u003c/sub\u003eO, organic mattter, total nitrogen (TN), P\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, and K\u003csub\u003e2\u003c/sub\u003eO at 55%, 34.5 g kg\u003csup\u003e-1\u003c/sup\u003e, 16.5 g kg\u003csup\u003e-1\u003c/sup\u003e, 23.6 g kg\u003csup\u003e-1\u003c/sup\u003e, and 15.8 g kg\u003csup\u003e-1\u003c/sup\u003e, respectively. The fertilization treatment involved a combination of chemical fertilizers and organic fertilizer. \u003cem\u003eScutellaria baicalensis\u003c/em\u003e seeds were randomly sown in experimental plots of 60 square meters, with each plot containing 1300 plants. The row spacing was 30 cm, and the plant spacing was 15 cm.\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003eSoil sampling\u003c/h3\u003e\n\u003cp\u003eAt the flowering stage, fruiting stage, and wilting stage in 2024, fifteen \u003cem\u003eScutellaria baicalensis\u003c/em\u003e plants were randomly selected from each replicate plot. The \u003cem\u003eScutellaria baicalensis\u003c/em\u003e roots with soil were shaken to remove loose soil, and the soil adhering to the root surface was brushed off with a soft brush, which was considered rhizosphere soil. The rhizosphere soil from fifteen \u003cem\u003eScutellaria baicalensis\u003c/em\u003e roots was mixed to form one soil sample for each treatment. Each soil sample is divided into three portions, which are used for chemical composition analysis, enzyme activity determination, and microbial diversity analysis respectively.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMeasurement of\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e \u003cstrong\u003egrowth indicators\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter collecting rhizosphere soil samples at the fruiting stage, the \u003cem\u003eScutellaria baicalensis\u003c/em\u003e plants were used to determine shoot fresh weight (SFW), root fresh weight (RFW), shoot dry weight (SDW) and root dry weight (RDW). Additionally, dry/fresh ratio (D/F) and root/shoot ratio (R/S), SDW/SFW and RDW/RFW were calculated. R/S\u0026thinsp;=\u0026thinsp;RDW/SDW.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetection of active components in\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e \u003cstrong\u003eroot\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHigh-performance liquid chromatography (HPLC) was used to determine baicalin, wogonoside, baicalein, and wogonin. Chromatographic column: kromasil eternity-5-C18 (250 mm\u0026times;4.6 mm); mobile phase: acetonitrile solution (A)-0.4% acetic acid solution (B), gradient elution; column temperature: 30\u0026deg;C; detection wavelength: 280 nm; flow rate: 1.0 mL/min.\u003c/p\u003e\n\u003cp\u003e1.6 Measurement of soil nutrients, microbial biomass carbon, nitrogen ratio, and enzyme activity\u003c/p\u003e\n\u003cp\u003eThe contents of soil organic carbon (SOC), total nitrogen (TN), alkali-hydrolyzable nitrogen (AN), total phosphorus (TP), available phosphorus (AP), total potassium (TK), and available potassium (AK) were determined using the method of Bao (Bao \u003cspan class=\"CitationRef\"\u003e2000\u003c/span\u003e). The contents of soil microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN) were also measured. The activities of urease (UR), alkaline phosphatase (PP), and sucrase (SUC) were determined using the method of Guan (Guan \u003cspan class=\"CitationRef\"\u003e1986\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eSoil DNA extraction, Illumina sequencing, and data processing\u003c/h3\u003e\n\u003cp\u003eUsing the Soil DNA Extraction Kit from Omega Bio-Tec (Norcross, GA, US), microbial genomic DNA was extracted from soil samples. The bacterial 16S rDNA V3-V4 region was amplified using BacF:ACTCCTACGGGAGGCAGCA and Bac R:GGACTACHVGGGTWTCTAAT; the fungal ITS region was amplified using ITSF (5\u0026prime; GGAAGTAAAAGTCGTAACAAGG-3\u0026prime;)/ITSR (5\u0026prime;-GCTGCG TTCTTCATCGATGC-3\u0026prime;). PCR conditions were 98\u0026deg;C for 5 min, 98\u0026deg;C for 30 s, 53\u0026deg;C for 30 s, 25 cycles, 72\u0026deg;C for 45 s, 72\u0026deg;C for 5 min, and 12\u0026deg;C. The amplification results were subjected to 2% agarose gel electrophoresis, and the target fragments were excised and recovered using the Axygen Gel Extraction Kit. The DADA2 Method (Benjamin et al., 2016) was used to process the raw sequencing data, including primer removal, quality filtering to exclude low-quality sequences, denoising to correct noisy reads, automatic sequence merging for paired-end data, chimera detection to remove chimeric sequences, and deduplication. The deduplicated sequences generated after processing are called Amplicon Sequence Variants. Meanwhile, a feature table recording the abundance of these sequences in each sample is generated. These are the core data for subsequent microbiome analysis. Sequencing data accession numbers for bacteria and fungi in NCBI are PRJNA1303951 and PRJNA1303954, respectively.\u003c/p\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003eData analysis\u003c/h2\u003e\n \u003cp\u003eQime software was used to calculate Chao, Ace, and Shannon diversity index. R software was used to generate Venn Diagrams. Canoco 5.0 was used for Redundancy Analysis (RDA). Prism software was used to create bar charts. SPSS 16.0 software was used for One-Way ANOVA to analyze differences. Each experiment was repeated three times, and the measured values were expressed as Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;Standard Error (SE). Different letters show significant difference at P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 using Waller Duncan test.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eChanges in soil physicochemical properties of\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e \u003cstrong\u003eat three different growth stages\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eChanges in the physicochemical properties of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil at three different growth stages are shown in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. Compared with the flowering stage, the contents of TK, AN, SOC,MBC, MBN, and the C/N ratio increased significantly by 6.53%, 23.86%, 31.93%, 49.01%, 31.01%, 19.52% at the fruiting stage and by 17.34%, 52.32%, 42.32%, 62.20%, 73.39%, 29.94% at the withering stage, with the order being withering stage\u0026thinsp;\u0026gt;\u0026thinsp;fruiting stage\u0026thinsp;\u0026gt;\u0026thinsp;flowering stage for all of them. The contents of TP and AP significantly increased by 38.83%, 50.68% and 27.18%, 36.59% at the fruiting stage and wilting stage compared with the flowering stage, respectively, with the order being fruiting stage\u0026thinsp;\u0026gt;\u0026thinsp;wilting stage\u0026thinsp;\u0026gt;\u0026thinsp;flowering stage. TP content was higher at the fruiting stage and wilting stage compared with the flowering stage, with little difference between the fruiting stage and wilting stage. Compared with the flowering stage, the gradual increase in contents of TN, TP, TK, AN, AP, AK, SOC, MBC, MBN, and the C/N ratio at the fruiting stage and wilting stage may be related to the gradual release of fertilizer effects after the application of organic fertilizer. pH value was the highest at the flowering stage (7.8), followed by the fruiting stage (7.5), and the lowest at the wilting stage (7.2). The gradual decrease in pH value may be related to the accumulation of rhizosphere exudates from \u003cem\u003eScutellaria baicalensis\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eChanges in plant mass accumulation of\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e \u003cstrong\u003eat different growth stages\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSoil physicochemical properties of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e at different growth stages\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003epH\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTN g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTP g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003cp\u003e(g/kg)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTK g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003cp\u003e(g/kg)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eAN mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eAP mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlowering stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.59a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e33.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFruiting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e53.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.99b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12c\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWilting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e66.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56b\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAK mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSOC g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMBC mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMBN mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC/N\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e--\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlowering stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e138.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.04b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e159.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e41.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e--\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFruiting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e151.87\u0026thinsp;\u0026plusmn;\u0026thinsp;1.10c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e236.93\u0026thinsp;\u0026plusmn;\u0026thinsp;1.11b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e54.50\u0026thinsp;\u0026plusmn;\u0026thinsp;1.28b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e--\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWilting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e134.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e257.90\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e72.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e--\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003eValues are given as the means\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;3); Values within a row followed by different lowercase letters are\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003esignificantly different (n\u0026thinsp;=\u0026thinsp;3, LSD, P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe results are shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. Compared with the flowering stage, RFW and RDW increased significantly by 60.00% and 50.99% respectively at the fruiting stage, and by 97.43% and 82.12% respectively at the withering stage. Compared with the flowering stage, SFW and RFW increased by 12.48% and 20.70% at the fruiting stage, and decreased by 41.767% and 32.26% at the wilting stage, respectively. Compared with the flowering stage, the R/S and SDW/SFW increased significantly by 25.09%, 7.31% at the fruiting stage and 168.86%, 16.32% at the withering stage, respectively; the RDW/RFW significantly decreased by 5.63% and 7.76% at the fruiting stage and wilting stage, respectively.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAccumulation of active components in\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e \u003cstrong\u003eat different growth stages\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAs shown in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, compared with the flowering stage, the contents of four components at the fruiting stage and wilting stage increased significantly, with the following percentage increases at the fruiting stage baicalin (83.87%), wogonoside (82.00%), baicalein (48.15%), and wogonin (61.54%) and at the wilting stage baicalin (114.52%), wogonoside (84.54%), baicalein (204.94%), and wogonin (210.26%).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffects of fertilization on soil enzyme activity of\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe enzyme activities in the rhizosphere soil at the growth and development of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. Compared with the flowering stage, the activities of UR, PP, and SUC significantly increased by 100.67%, 29.30%, and 35.71% at the fruiting stage, and significantly increased significantly increased by 80.43%, 68.23%, and 59.07% at the wilting stage.This indicates that soil enzyme activity gradually increases with the growth stage and the gradual increase in rhizosphere soil nutrients\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eContents of active components in roots at different growth stages\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eBaicalin\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWogonoside\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eBaicalein\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWogonin\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlowering stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFruiting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWilting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01c\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eValues are given as the means\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;3); Values within a row followed by different lowercase letters are\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003esignificantly different (n\u0026thinsp;=\u0026thinsp;3, LSD, P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eEffects of fertilization on soil microbial diversity of\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe used high-throughput sequencing technology to analyze the differences in bacterial and fungal community composition in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere at the flowering stage, fruiting stage, and wilting stage. Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e shows that the mean values of high-quality sequences of bacteria at the flowering stage, fruiting stage, and wilting stage were 53113, 55521, and 59101, respectively. Compared with the flowering stage, the high-quality sequences and Shannon index significantly increased by 4.53%, 3.06% (at the fruiting stage) and 11.27%, 4.20% (at the wilting stage), respectively. Additionally, the Chao index significantly increased by 9.85% at the wilting stage compared with the flowering stage. For fungi, the values of high-quality sequences were 46564, 43068, and 42507, respectively. Compared with the flowering stage, the Chao index and Simpson index significantly increased by 7.95% and 4.54% at the fruiting stage, and by 8.46% and 6.10% at the wilting stage. Shannon index significantly increased by 9.09% at the wilting stage compared with the flowering stage. High-quality sequences significantly decreased by 7.51% and 8.71% at the fruiting stage and wilting stage compared with the flowering stage.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSequence numbers and diversity indices of rhizosphere soil of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e at different growth stages\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eChao\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eShannon\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSimpson\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003ehigh-quality sequences\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlowering stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3580.53\u0026thinsp;\u0026plusmn;\u0026thinsp;80.21a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9982\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0001a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e53113\u0026thinsp;\u0026plusmn;\u0026thinsp;135a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFruiting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3601.11\u0026thinsp;\u0026plusmn;\u0026thinsp;16.12a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9987\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e55521\u0026thinsp;\u0026plusmn;\u0026thinsp;89b\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWilting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3856.21\u0026thinsp;\u0026plusmn;\u0026thinsp;38.12c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9999\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0002b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e59101\u0026thinsp;\u0026plusmn;\u0026thinsp;98c\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eChao\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eShannon\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSimpson\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003ehigh-quality sequences\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlowering stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e288.67\u0026thinsp;\u0026plusmn;\u0026thinsp;6.61b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9017\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0114a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e46564\u0026thinsp;\u0026plusmn;\u0026thinsp;70.15c\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFruiting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e265.72\u0026thinsp;\u0026plusmn;\u0026thinsp;1.96a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9426\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0027b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e43068\u0026thinsp;\u0026plusmn;\u0026thinsp;48.50b\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWilting stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e313.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9567\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0026c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e42507\u0026thinsp;\u0026plusmn;\u0026thinsp;67.28a\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003eValues are given as the means\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;3); Values within a row followed by different lowercase letters are\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003esignificantly different (n\u0026thinsp;=\u0026thinsp;3, LSD, P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eChanges in bacterial composition at the phylum level in rhizosphere soil of\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e \u003cstrong\u003eat different growth stages\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn each sample, genera with relative abundance exceeding 0.5% were analyzed at the phylum level (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). At the growth stage of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e, the relative abundance of main bacterial communities in rhizosphere soil (including \u003cem\u003eActinobacteriota\u003c/em\u003e, \u003cem\u003eBacteroidota\u003c/em\u003e, \u003cem\u003eProteobacteria\u003c/em\u003e, and \u003cem\u003eChloroflexi\u003c/em\u003e significantly increased by 15.44%, 11.59%, 26.01%, and 10.88% respectively at the fruiting stage, and by 22.49%, 31.72%, 13.52%, and 22.76% respectively at the wilting stage, compared with the flowering stage. The relative abundance of \u003cem\u003eAcidobacteria\u003c/em\u003e, \u003cem\u003eFirmicutes\u003c/em\u003e, \u003cem\u003eVerrucomicrobiot\u003c/em\u003ea, and \u003cem\u003ePlanctomycetota\u003c/em\u003e significantly decreased by 8.43%, 13.36%, 35.75%, and 26.82% at the fruiting stage and 16.65%, 19.82%, 57.56%, and 50.85% at the wilting stage compared with the flowering stage. The relative abundance of \u003cem\u003eGemmatimonadota\u003c/em\u003e significantly increased by 15.59% at the fruiting stage compared with the flowering stage, with little difference between the wilting stage and the flowering stage.\u003c/p\u003e\n\u003cp\u003eAt different growth stages of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e, the relative abundance of main fungal communities in rhizosphere soil, such as \u003cem\u003eAscomycota\u003c/em\u003e, was 93.67%, 95.52%, and 91.41% at the flowering stage, fruiting stage, and wilting stage, respectively. The relative abundance of \u003cem\u003eMortierellomycota\u003c/em\u003e, \u003cem\u003eAphelidiomycota\u003c/em\u003e, and \u003cem\u003eChytridiomycota\u003c/em\u003e significantly decreased by 13.46%, 52.38%, and 25.66% at the fruiting stage and 49.20%, 63.18%, and 61.03% at the wilting stage compared with the flowering stage. The relative abundance of \u003cem\u003eBasidiomycota\u003c/em\u003e significantly increased by 50.41% at the fruiting stage and by 207.91% at the wilting stage compared with the flowering stage.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eChanges in bacterial composition at the genus level in\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e \u003cstrong\u003erhizosphere soil at different growth stage\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn each sample, genera with relative abundance exceeding 0.5% were analyzed at the genus level (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Among 30 bacterial genera, \u003cem\u003eSphingomonas\u003c/em\u003e had the highest relative abundance, with values of 1.52%, 2.34%, and 2.07% at the flowering stage, fruiting stage, and wilting stage, respectively. Compared with the flowering stage, the relative abundance of 9 bacterial genera (\u003cem\u003eHaliangium\u003c/em\u003e, \u003cem\u003eBlastococcus\u003c/em\u003e, \u003cem\u003eSolirubrobacter\u003c/em\u003e, \u003cem\u003eStreptomyces\u003c/em\u003e, \u003cem\u003eCellulomonas\u003c/em\u003e, \u003cem\u003eRubrobacter\u003c/em\u003e, \u003cem\u003eEnsifer\u003c/em\u003e, \u003cem\u003eGeorgenia\u003c/em\u003e, and \u003cem\u003eIamia\u003c/em\u003e) exhibited a significant increase, specifically rising by 22.74\u0026ndash;73.58% at the fruiting stage and by 9.46\u0026ndash;42.43% at the wilting stage. The relative abundance of these 9 genera increased from the flowering stage to the fruiting stage and decreased from the fruiting stage to the wilting stage, with the fruiting stage being higher than the flowering stage. The relative abundance of \u003cem\u003eBradyrhizobium\u003c/em\u003e increased by 50.23% at the fruiting stage compared with the flowering stage, with little difference between the wilting stage and the flowering stage. Compared with the flowering stage, the relative abundance of \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003eNocardioides\u003c/em\u003e, \u003cem\u003eGeodermatophilus\u003c/em\u003e, \u003cem\u003eMycobacterium\u003c/em\u003e, \u003cem\u003eBryobacter\u003c/em\u003e, \u003cem\u003ePseudomonas\u003c/em\u003e, \u003cem\u003eSkermanella\u003c/em\u003e, \u003cem\u003eAgromyces\u003c/em\u003e, and \u003cem\u003eDevosia\u003c/em\u003e significantly decreased by 8.08%~31.32% at the fruiting stage and by 23.40%~57.59% at the wilting stage. Compared with the flowering stage, the relative abundance of \u003cem\u003eGemmatimonas\u003c/em\u003e, \u003cem\u003eLysobacter\u003c/em\u003e, \u003cem\u003eRamlibacter\u003c/em\u003e, \u003cem\u003ePedomicrobium\u003c/em\u003e, \u003cem\u003eRhodomicrobium\u003c/em\u003e, \u003cem\u003eAeromicrobium\u003c/em\u003e, \u003cem\u003ePhenylobacterium\u003c/em\u003e, \u003cem\u003eOpitutus\u003c/em\u003e, \u003cem\u003eArenimonas\u003c/em\u003e, and \u003cem\u003eCandidatus Solibacter\u003c/em\u003e significantly increased by 14.76%~48.60% at the fruiting stage and by 16.34%~175.98% at the wilting stage. The relative abundance of these 10 genera gradually increased at the flowering stage, fruiting stage, and wilting stage.\u003c/p\u003e\n\u003cp\u003eAmong 30 fungal genera, \u003cem\u003eMycochlamy\u003c/em\u003es had the highest relative abundance, with values of 4.41%, 6.78%, and 7.82% at the flowering stage, fruiting stage, and wilting stage, respectively. Compared with the flowering stage, the relative abundance of \u003cem\u003eTrichocladium\u003c/em\u003e, \u003cem\u003eFusarium\u003c/em\u003e, \u003cem\u003ePlectosphaerella\u003c/em\u003e, \u003cem\u003eCladosporium\u003c/em\u003e, \u003cem\u003eScopulariopsis\u003c/em\u003e, \u003cem\u003eSodiomyces\u003c/em\u003e, \u003cem\u003eMortierella\u003c/em\u003e, \u003cem\u003ePodospora\u003c/em\u003e, \u003cem\u003eMetarhizium\u003c/em\u003e, and \u003cem\u003eMyrmecridium\u003c/em\u003e significantly increased by 24.22%~195.57% at the fruiting stage and by 43.66%~418.83% at the wilting stage.The relative abundance of these 11 genera gradually increased in rhizosphere soil during the growth stage of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e. The relative abundance of \u003cem\u003eAlternaria\u003c/em\u003e significantly increased by 90.67% at the wilting stage compared with the flowering stage and fruiting stage, with little difference between the flowering stage and fruiting stage. Compared with the flowering stage, the relative abundance of \u003cem\u003ePreussia\u003c/em\u003e, \u003cem\u003eArthrographis\u003c/em\u003e, \u003cem\u003eEremomyces\u003c/em\u003e, \u003cem\u003eCorynascella\u003c/em\u003e, \u003cem\u003eZopfiella\u003c/em\u003e, \u003cem\u003eBisifusarium\u003c/em\u003e, \u003cem\u003ePenicillium\u003c/em\u003e, \u003cem\u003eBeauveria\u003c/em\u003e, \u003cem\u003eSolicoccozyma\u003c/em\u003e, and significantly decreased by 4.05%~82.83% at the fruiting stage and by 24.88%~90.58% at the wilting stage. The relative abundance of these 10 genera gradually decreased in rhizosphere soil at the growth stage of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e. The relative abundance of \u003cem\u003eBotryotrichum\u003c/em\u003e, \u003cem\u003ePseudogymnoascus\u003c/em\u003e, \u003cem\u003eAspergillus\u003c/em\u003e, \u003cem\u003eXenodidymella\u003c/em\u003e, and \u003cem\u003eSubramaniula\u003c/em\u003e significantly increased by 28.55%~234.19% at the fruiting stage compared with the flowering stage and significantly decreased by 14.03%~47.47% at the wilting stage compared with the flowering stage. The relative abundance of these 6 genera showed a trend of first increasing and then decreasing at the three growth stages of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e, with the relative abundance at the wilting stage being lower than that of the flowering stage. The relative abundance of \u003cem\u003eAcaulium\u003c/em\u003e and \u003cem\u003eStolonocarpus\u003c/em\u003e significantly decreased by 66.07% and 75.90% at the fruiting stage compared with the flowering stage, with little difference between the flowering stage and wilting stage.\u003c/p\u003e\n\u003cp\u003eCompared with the flowering stage, the relative abundance of \u003cem\u003eWardomyces\u003c/em\u003e and \u003cem\u003ePseudallescheria\u003c/em\u003e significantly increased by 43.47% and 6.53% at the fruiting stage, and by 262.99% and 64.06% at the wilting stage, respectively.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRelationship between rhizosphere soil nutrients, microbial groups, and enzyme activity of\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eRDA revealed the relationship between soil chemical properties, microbial community, and enzyme activity, with soil properties as environmental factors and microbial groups as variables (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003ea, b, c, d). Axis 1 and Axis 2 explain 75.48% and 20.29% of the total variation between the community composition at the bacterial phyla level and soil properties (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003ea). The contents of AN, AP, AK, SOC, MBC, MBN, and C/N ratio were significantly positively correlated with the relative abundance of \u003cem\u003eBacteroidetes\u003c/em\u003e, \u003cem\u003eGemmatimonadetes\u003c/em\u003e, \u003cem\u003eProteobacteria\u003c/em\u003e, \u003cem\u003eChloroflexi\u003c/em\u003e, \u003cem\u003eActinobacteria\u003c/em\u003e, and \u003cem\u003eCyanobacteria\u003c/em\u003e; The pH value was significantly positively correlated with the relative abundance of \u003cem\u003eVerrucomicrobia\u003c/em\u003e, \u003cem\u003eFirmicutes\u003c/em\u003e, \u003cem\u003eAcidobacteria\u003c/em\u003e, \u003cem\u003eNitrospirae\u003c/em\u003e, \u003cem\u003ePlanctomycetes\u003c/em\u003e, and \u003cem\u003eArmatimonadetes\u003c/em\u003e. The most significant factor affecting bacteria at the phyla level was AP (F\u0026thinsp;=\u0026thinsp;19.8, P\u0026thinsp;=\u0026thinsp;0.002), followed by MBC (F\u0026thinsp;=\u0026thinsp;14.9, P\u0026thinsp;=\u0026thinsp;0.014). Axis 1 and Axis 2 explain 86.70% and 4.84% of the total variation between the community composition at the bacterial genus level and soil properties (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eb).The contents of AN, AP, AK, MBN, MBC, and C/N ratio were positively correlated with the relative abundance of \u003cem\u003eGemmatimonas\u003c/em\u003e, \u003cem\u003ePedomicrobium\u003c/em\u003e, \u003cem\u003eAeromicrobium\u003c/em\u003e, \u003cem\u003eArenimonas\u003c/em\u003e, \u003cem\u003eLysobacter\u003c/em\u003e, \u003cem\u003eRamlibacter\u003c/em\u003e, \u003cem\u003eRhodomicrobium\u003c/em\u003e, \u003cem\u003ePhenylobacterium\u003c/em\u003e, \u003cem\u003eOpitutus\u003c/em\u003e, and \u003cem\u003eCandidatus Solibacter\u003c/em\u003e. The most significant factor affecting bacteria at the genus level was AN (F\u0026thinsp;=\u0026thinsp;35.8, P\u0026thinsp;=\u0026thinsp;0.002), followed by MBN (F\u0026thinsp;=\u0026thinsp;30.0, P\u0026thinsp;=\u0026thinsp;0.004). Axis 1 and Axis 2 explain 83.45%和15.02% of the total variation between the community composition at the fungal phyla level and soil properties (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003ec). RDA showed that the pH value and AK content were positively correlated with the relative abundance of \u003cem\u003eAphelidiomycota\u003c/em\u003e, \u003cem\u003eMortierellomycota\u003c/em\u003e, \u003cem\u003eAscomycota\u003c/em\u003e, and \u003cem\u003eChytridiomycota\u003c/em\u003e; the relative abundance of \u003cem\u003eBasidiomycota\u003c/em\u003e was positively correlated with the contents of AN, AP, SOC, MBC, MBN, and C/N ratio. The most significant factor affecting fungi at the phyla level was MB (F\u0026thinsp;=\u0026thinsp;9.7, P\u0026thinsp;=\u0026thinsp;0.018), followed by AN (F\u0026thinsp;=\u0026thinsp;9.0, P\u0026thinsp;=\u0026thinsp;0.018). Axis 1 and Axis 2 explain 97.28%和2.34% of the total variation between the community composition at the fungal phyla level and soil properties (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003ed).RDA showed that the contents of AN, AP, AK, MBN, MBC, and C/N ratio were positively correlated with the relative abundance of \u003cem\u003eMycochlamys\u003c/em\u003e, \u003cem\u003eTrichocladium\u003c/em\u003e, \u003cem\u003eFusarium\u003c/em\u003e, \u003cem\u003ePlectosphaerella\u003c/em\u003e, \u003cem\u003eCladosporium\u003c/em\u003e, \u003cem\u003eAlternaria\u003c/em\u003e, \u003cem\u003eScopulariopsis\u003c/em\u003e, \u003cem\u003eSodiomyces\u003c/em\u003e, \u003cem\u003eMortierella\u003c/em\u003e, \u003cem\u003ePodospora\u003c/em\u003e, \u003cem\u003eMetarhizium\u003c/em\u003e, and \u003cem\u003eMyrmecridium\u003c/em\u003e. The most significant factor affecting fungi at the genus level was MBN (F\u0026thinsp;=\u0026thinsp;117, P\u0026thinsp;=\u0026thinsp;0.004), followed by AN (F\u0026thinsp;=\u0026thinsp;152, P\u0026thinsp;=\u0026thinsp;0.004). Axis 1 and Axis 2 explain 95.23%和4.63% of soil enzyme activities and soil properties (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003ee). PP activity was positively correlated with the contents of AN, AP, MBC, MBN, SOC, and C/N ratio; UR activity was positively correlated with the contents of AN, AP, AK, MBC, MBN, SOC, and C/N ratio; SUC activity was positively correlated with the contents of AN, AP, MBC, MBN, SOC, and C/N ratio. The activities of the three enzymes significantly influenced soil physicochemical properties.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRelationship between effective components and growth condition of\u003c/strong\u003e \u003cstrong\u003eScutellaria baicalensis\u003c/strong\u003e \u003cstrong\u003eroot and microbial community.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFigures 7a and 7b are correlation heatmaps of bacterial and fungal communities with plant effective components and growth traits, respectively. The heatmaps visually present the Pearson correlation between plant indicators and microbial communities through color. Figure a shows that baicalin, wogonoside, baicalein, wogonin, SDW/SDF, and R/S were significantly positively correlated with 10 bacterial genera such as \u003cem\u003eGemmatimonas\u003c/em\u003e and \u003cem\u003ePedobacter\u003c/em\u003e, while negatively correlated with 9 bacterial genera such as \u003cem\u003eBacillus\u003c/em\u003e and \u003cem\u003eNocardioides\u003c/em\u003e; the RDW/RFW showed the opposite trend. Figure b shows that baicalin, wogonoside, baicalein, wogonin, SDW/SFW, and R/S were significantly positively correlated with 12 fungal genera such as \u003cem\u003eMycochlamys\u003c/em\u003e and \u003cem\u003eTrichocladium\u003c/em\u003e, while negatively correlated with 9 fungal genera such as \u003cem\u003ePreussia\u003c/em\u003e and \u003cem\u003eArthrographis\u003c/em\u003e; the RDW/RFW showed the opposite trend.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eDuring the cultivation of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e, soil nutrients play an important role in promoting the formation of yield and quality. Studies have shown that nutrient cycling is closely related to soil bacterial diversity and function, especially the changes in soil nutrients at different growth stages after \u003cem\u003eScutellaria baicalensis\u003c/em\u003e planting, which affect the bacterial diversity in rhizosphere soil. Meanwhile, soil enzyme activity plays an important role in soil nutrient transformation. This study found that from the flowering stage to the wilting stage, urease activity in the soil gradually increased, and AN content correspondingly increased, indicating that the increase in urease activity promoted the decomposition of organic nitrogen in the soil, increasing the AN content (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). This is consistent with the findings of Wu in \u003cem\u003eFructus aurantii\u003c/em\u003e (Wu et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). At the wilting stage, soil PP activity was higher than in the fruiting stage, while the total AP content in the soil was lower (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). This may be because the demand for phosphorus element increases with the growth of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e, reducing the AP content in the soil. Previous studies on \u003cem\u003eRehmannia glutinosa\u003c/em\u003e found that appropriate application of phosphate fertilizer increased the phosphorus content in \u003cem\u003eRehmannia glutinosa\u003c/em\u003e roots (Gao et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), supporting our result.\u003c/p\u003e\u003cp\u003eIn our previous experiments on different fertilization measures for \u003cem\u003eScutellaria baicalensis\u003c/em\u003e, we found that increasing organic fertilizer could improve the content of soil organic matter. Therefore, this experiment adopted the measure of combining organic fertilizer and inorganic fertilizer, which is consistent with the findings of Sun in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e fertilization experiments (Sun et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Additionally, changes in soil nutrients can alter the bacterial diversity and function in rhizosphere soil. In this study, compared with the flowering stage, the Shannon index and Chao index of the soil increased at the fruiting and wilting stages (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e), indicating that soil organic matter facilitates the formation of soil bacterial diversity.\u003c/p\u003e\u003cp\u003eTo confirm our hypothesis, we further analyzed the functions of bacteria and fungi at the phyla level during different growth stages of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e. From the flowering stage to the withering stage, the relative abundance of \u003cem\u003eActinobacteria\u003c/em\u003e and \u003cem\u003eBacteroidetes\u003c/em\u003e gradually increased, and the contents of AN, SOC, MBC, MBN, and C/N ratio in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil showed a gradual increasing trend (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The reasons are, firstly, the continuous release of fertilizer efficiency from organic fertilizer in the soil during the growth and development of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e promotes the reproduction of nutrient-rich bacteria such as \u003cem\u003eActinobacteria\u003c/em\u003e and \u003cem\u003eBacteroidetes\u003c/em\u003e, and \u003cem\u003eActinobacteria\u003c/em\u003e contributes to the degradation and utilization of organic matter; secondly, from the flowering stage to the fruiting stage, the vigorous growth of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e roots produces a large amount of rhizosphere exudates, promoting the formation of organic matter in the soil, and the rich organic matter benefits the growth of \u003cem\u003eActinobacteria\u003c/em\u003e. This study found that with the growth and development of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e, the abundances of \u003cem\u003eProteobacteria\u003c/em\u003e and \u003cem\u003eChloroflexi\u003c/em\u003e in the soil both showed a trend of first increasing and then decreasing (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). This variation pattern was consistent with the content dynamics of AK and AP in the soil, and also aligned with the conclusion from previous research that \u003cem\u003eProteobacteria\u003c/em\u003e are eutrophic bacteria (Fierer et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). However, this trend was not entirely consistent with the existing finding that \u003cem\u003eChloroflexi\u003c/em\u003e are oligotrophic bacteria (Xian et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). We speculate that this may vary depending on plant species. At the genus level, the study found that \u003cem\u003eBacillus\u003c/em\u003e spp. in the rhizosphere participates in the cycling of proteins and cellulose, and \u003cem\u003eBacillus velezensis\u003c/em\u003e YH-18 and \u003cem\u003eBacillus velezensis\u003c/em\u003e YH-20 can promote the growth of stems and leaves of peach plants and the cultivation of high-quality tree species (Shi et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). \u003cem\u003eGeodermatophilus obcurus\u003c/em\u003e, isolated from dolomite marble, can tolerate harsh environments such as dryness, mitomycin C, hydrogen peroxide, ionizing radiation, and ultraviolet radiation (Montero-Calasanz et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). \u003cem\u003eBryobacter\u003c/em\u003e spp. can decompose and utilize polysaccharides, various sugars, and organic acids in carbon cycling (Li et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). \u003cem\u003eNocardioides\u003c/em\u003e spp. not only has phosphorus-solubilizing functions but also promotes the degradation of various hard-to-degrade organic compounds such as aromatic compounds, hydrocarbons, halogenated alkanes, nitrogen-containing heterocyclic compounds, and polymeric polyesters (Wang et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2022b\u003c/span\u003e); \u003cem\u003eSphingomonas\u003c/em\u003e spp. can degrade 3-(4-isopropylphenyl)-1,1-dimethylurea in soils with pH above 7.0 (Bending et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). \u003cem\u003eStreptomyces\u003c/em\u003e spp. participates in the decomposition of organic matter and can promote the activation of plant disease resistance mechanisms; compared with normal nitrogen fertilizer application, reducing nitrogen fertilizer by 50% combined with \u003cem\u003eStreptomyces\u003c/em\u003e sp. NGB-Act4 and NGB-Act6 significantly promoted wheat growth and increased yield (Youseif et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023\u003c/span\u003e); further demonstrating the growth-promoting effect of \u003cem\u003eStreptomyces\u003c/em\u003e spp. on plants. Our study found that \u003cem\u003eStreptomyces\u003c/em\u003e spp. first increased and then decreased (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e), indicating that this genus played an active role during the vigorous growth stage of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e. \u003cem\u003eBlastococcus\u003c/em\u003e spp. is widely present in soils with extreme cold, extreme heat, salinization, toxic amendments, and significant heavy metal pollution, and can be used for soil improvement (Sbissi et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2025\u003c/span\u003e); \u003cem\u003eHaliangium\u003c/em\u003e can be used as a plant bactericide (Kumar et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). \u003cem\u003eOpitutus\u003c/em\u003e has nitrogen fixation and nitrogen preservation functions (Hu et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). \u003cem\u003eLysobacter enzymogenes\u003c/em\u003e is a non-pathogenic strain that inhibits various crop fungal diseases by synthesizing antifungal factors (Zhao et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). \u003cem\u003eGemmatimonas\u003c/em\u003e spp. participates in the reduction of N₂O, reducing nitrogen loss in the form of N₂O, allowing more nitrogen to remain in the soil for crop absorption, thereby improving nitrogen fertilizer utilization efficiency and reducing agricultural production costs (Oshiki et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The study of bacterial functions shows that the above 11 genera are related to nutrient cycling, organic matter degradation, or antagonism against soil pathogens. 4 genera had the highest abundance at the flowering stage, 4 genera had the highest relative abundance at the fruiting stage, and 3 genera had the highest relative abundance at the wilting stage. This indicates that the communities and abundance of beneficial bacteria in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere at the three different growth stages show regular changes. Correlation analysis showed that \u003cem\u003eOpitutus\u003c/em\u003e, \u003cem\u003eLysobacter\u003c/em\u003e, and \u003cem\u003eGemmatimonas\u003c/em\u003e were significantly positively correlated with the accumulation of effective components (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e), further supporting our results.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eResearch on fungal functions has found that \u003cem\u003eAscomycota\u003c/em\u003e has higher abundance in soils with lower organic matter than in soils with higher organic matter. This phylum can participate in the formation of soil aggregates and serves as an important source of various toxins. (Li et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Fu et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e); \u003cem\u003eBasidiomycota\u003c/em\u003e is saprophytic fungi suitable for survival in environments rich in organic matter (Fu et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The results of this study showed that compared with the flowering stage, the relative abundance of \u003cem\u003eAscomycota\u003c/em\u003e increased at the fruiting stage and decreased at the wilting stage, while the relative abundance of \u003cem\u003eBasidiomycot\u003c/em\u003ea gradually increased at the fruiting and wilting stages; this is related to the gradual increase in soil organic matter with the growth and development of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e. \u003cem\u003eArthrographis\u003c/em\u003e spp. has the ability to produce cellulase and is commonly found in compost (Eida et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Okeke et al. 1993). Some species of \u003cem\u003ePenicillium\u003c/em\u003e spp. can produce soluble phosphorus, iron carriers, and plant hormones (such as indole-3-acetic acid and gibberellin), which play an important role in plant health (Park et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). \u003cem\u003ePreussia\u003c/em\u003e spp. can produce plant growth hormones to promote plant growth (Al-Hosni et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). \u003cem\u003eAspergillus brunneoviolaceus\u003c/em\u003e HZ23 is a plant growth-promoting bacterium that changes the physicochemical properties of rhizosphere soil of pak choi and can be used as an amendment for newly reclaimed soil (Li et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). \u003cem\u003ePseudogymnoascus\u003c/em\u003e spp. is the most abundant fungal genus in the rhizosphere, and its members are usually involved in cellulose degradation (Sigler et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). \u003cem\u003ePseudallescheria\u003c/em\u003e spp. is a potential pathogen found in soil and on humans and animals (Rainer et al. 2020). \u003cem\u003eMycochlamys\u003c/em\u003e can participate in chitin degradation (Debode et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). A large amount of \u003cem\u003eMortierella alpina\u003c/em\u003e in the rhizosphere not only participates in the decomposition of plant residues and organic matter but also promotes plant growth by facilitating the biosynthesis of carotenoid derivatives and enhancing stress tolerance (Wani et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2017\u003c/span\u003e); some genera of \u003cem\u003eMortierella\u003c/em\u003e spp. can convert insoluble phosphorus in the soil into soluble phosphorus, improving plant phosphorus absorption capacity and promoting plant growth and development (Sang et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). \u003cem\u003eAlternaria\u003c/em\u003e spp. is a potential plant pathogenic fungus that can cause potato early blight, potato brown spot, and soybean black spot (Xu et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). \u003cem\u003eFusarium\u003c/em\u003e spp. can cause crop diseases such as potato dry rot and potato \u003cem\u003eFusarium wilt\u003c/em\u003e (Zhang et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The study of fungal functions shows that the above 10 genera are related to nutrient cycling, organic matter degradation, and plant pathogenicity. Two genera had the highest abundance at the flowering stage, 2 genera had the highest relative abundance at the fruiting stage, and 4 genera had the highest relative abundance at the wilting stage. The highest relative abundance of \u003cem\u003eMycochlamy\u003c/em\u003es, \u003cem\u003eMortierella\u003c/em\u003e, \u003cem\u003eAlternaria\u003c/em\u003e, and \u003cem\u003eFusarium\u003c/em\u003e at the wilting stage was significantly positively correlated with effective components such as baicalin (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e), further supporting our results.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe research results show that influenced by the growth stage, the soil physicochemical properties of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e at different growth stages exhibit various changes. The diversity of fungi and bacteria is the highest at the wilting stage compared with fruiting stage and wilting stage. The groups of functional microorganisms among fungi and bacteria exhibit distinct characteristics at different growth stages of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e. The association patterns between bacterial and fungal diversity and the accumulation of active components in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e root provide key data for in-depth analysis of the ecological interaction network between plants and microorganisms. This helps explore the bidirectional mechanisms of plant metabolites regulating microbial communities and microorganisms influencing plant growth and development. It also provides a theoretical basis for optimizing plant growth and enhancing the accumulation of secondary metabolites through microbial community regulation.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eLixia Xu and Minna Guo Wrote the main manuscript text and Meishan Yue and Chunyan Guo prepared figures 1-3. All authors reviewed the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e\u003cp\u003eThis research was supported by Scientific Research Project of Shanxi Provincial Administration of Traditional Chinese Medicine (2023ZYYB05; Scientific Research Fund for Doctors and Postdoctoral Researchers Coming to Shanxi (2025), Key Research and Development Program Project of Shanxi Province (201803D221011-3). We are very grateful to Shanghai Personal Biotechnology Co., Ltd for Illumina sequencing.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAl-Hosni K, Shahzad R, Khan AL, Imran QM, Al Harrasi AA, Al Rawahi AA, Asaf S, Kang S-M, Yun B-W, Lee I-J (2018) Preussia sp. BSL-10 producing nitric oxide, gibberellins, and indole acetic acid and improving rice plant growth. 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Plant Dis 103:1286\u0026ndash;1292\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Scutellaria baicalensis Georg, High-throughput sequencing, Soil microbial diversity, Soil physicochemical properties, Active components of roots","lastPublishedDoi":"10.21203/rs.3.rs-7476576/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7476576/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn this study, \u003cem\u003eScutellaria baicalensis\u003c/em\u003e Gerog plants at different growth stages and their rhizosphere soil were used as experimental materials. The microbial diversity, physicochemical properties, and enzyme activities of the rhizosphere soil were systematically determined, while the active component contents of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e shoots were analyzed. This research aims to provide a scientific basis for revealing the soil environment regulation mechanism underlying the accumulation of active components in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e root. It revealed the association mechanisms between rhizosphere soil micro-ecological characteristics at different growth stages and active component accumulation of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e root. The results showed that, compared with the flowering stage, the contents of total phosphorus (TP), alkali-hydrolyzable nitrogen (AN), soil organic carbon (SOC), microbial biomass carbon (MBC), and microbial biomass nitrogen (MBN), as well as the carbon/nitrogen (C/N) ratio, in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil gradually increased during the fruiting and wilting stages. In contrast, the soil pH showed an opposite trend. Among the other soil nutrients, the contents of TP, available phosphorus (AP), and available potassium (AK) were the highest at the fruiting stage, followed by the wilting stage, the lowest at the wilting stage. Additionally, the TN content was the lowest at the flowering stage, and it was higher at the fruiting and wilting stages. Compared with the flowering stage, the activities of urease, sucrase, and alkaline phosphatase in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil significantly increased at the fruiting and wilting stages. There are significant differences of bacterial community in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil at different growth stages (flowering stage, fruiting stage, and withering stage). The relative abundance of bacteria at the genus level in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil changes with the growth stages. The dominant genera in each stage is as follows: at the flowering stage, the dominant taxa are \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003eGeodermatophilus\u003c/em\u003e, and \u003cem\u003eBryobacte\u003c/em\u003e; at the fruiting stage, the dominant taxa are \u003cem\u003eStreptomyces\u003c/em\u003e, \u003cem\u003eSphingomonas\u003c/em\u003e, \u003cem\u003eBlastococcuss\u003c/em\u003e, \u003cem\u003eHaliangium\u003c/em\u003e, and \u003cem\u003eNocardioides\u003c/em\u003e; At thewithering stage, the dominant taxa are \u003cem\u003eGemmatimonas\u003c/em\u003e, \u003cem\u003eOpitutus\u003c/em\u003e, and \u003cem\u003eLysobacter\u003c/em\u003e. The relative abundance of fungi in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil at the genus level changes with the growth stages. The dominant genera in each stage are as follows: at the flowering stage, the genera with the highest relative abundance are \u003cem\u003eArthrographis\u003c/em\u003e, \u003cem\u003ePenicillium\u003c/em\u003e and \u003cem\u003ePreussia\u003c/em\u003e; at the fruiting stage, the genera with the highest relative abundance are \u003cem\u003ePseudogymnoascus\u003c/em\u003e, \u003cem\u003eAspergillus\u003c/em\u003e and \u003cem\u003ePseudallescheria\u003c/em\u003e; at the withering stage, the genera with the highest relative abundance are \u003cem\u003eMortierella\u003c/em\u003e, \u003cem\u003eAlternaria\u003c/em\u003e, \u003cem\u003eMycochlamys\u003c/em\u003e and \u003cem\u003eFusarium\u003c/em\u003e. Redundancy analysis indicated that AN and MBN were key environmental factors driving the differences of bacterial and fungal communities at the genus level in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e rhizosphere soil at different growth stages. Pearson correlation analysis further showed that the main active components of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e root, including baicalin, wogonoside, baicalein, and wogonin, were significantly positively correlated with the relative abundance of 10 bacterial genera such as \u003cem\u003eGemmatimonas\u003c/em\u003e, \u003cem\u003eOpitutus\u003c/em\u003e, and \u003cem\u003eLysobacter\u003c/em\u003e, as well as with the relative abundance of 12 fungal genera such as \u003cem\u003eMortierella\u003c/em\u003e, \u003cem\u003eAlternaria\u003c/em\u003e, and \u003cem\u003eMycochlamys\u003c/em\u003e. These results reveal that the rhizosphere microbial community plays an important role in the accumulation of root active components at different growth stages of \u003cem\u003eScutellaria baicalensis\u003c/em\u003e, providing a theoretical basis for the development of precision fertilization strategies and scientific management in \u003cem\u003eScutellaria baicalensis\u003c/em\u003e cultivation.\u003c/p\u003e","manuscriptTitle":"Analysis of the variation characteristics of Scutellaria baicalensis rhizosphere soil microorganisms at different growth stages and their relationship with the accumulation of active components","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-06 07:49:47","doi":"10.21203/rs.3.rs-7476576/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6ec26286-f03f-491a-a26e-c4b72639c9a6","owner":[],"postedDate":"September 6th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-12-03T13:23:53+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-06 07:49:47","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7476576","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7476576","identity":"rs-7476576","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}