Correlation analysis between ginsenoside content and rhizosphere soil microbial species in different forest types

Correlation analysis between ginsenoside content and rhizosphere soil microbial species in different forest types | 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 Correlation analysis between ginsenoside content and rhizosphere soil microbial species in different forest types Fengyu Pang, Xiaojia Ruan, Yugang Gao, Yan Zhao, Qun Liu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4487770/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 Inter-root microorganisms are major factors influencing the formation of high-quality herbs and promoting the accumulation of secondary metabolites, and this relationship has been well characterised in many supra-herbal medicinal herbs, but there is limited information on whether ginseng saponin accumulation is mediated by microorganisms in different forest types.Inter-root soil samples of ginseng and ginseng samples were collected from four different forest types (Theropencedrymion, Larch forest, Broad-leaved forest and Camphor pine forest) in the mountainous areas of Jingyu County, Jilin Province, China. The content of ginsenosides in the collected ginseng samples was determined by high performance liquid chromatography (HPLC). The results showed that the content of ginsenosides in Camphor pine forest was significantly higher than that in the other three forest types.The microorganisms in the soil samples were isolated and purified, and subsequently sequenced and analyzed by high-throughput sequencing methods, and a total of seven bacterial species were isolated and identified in the inter-root soil of ginseng from four different forest types. In broad-leaved forests (BF) and larch forests (LF), Bacillus megaterium is the most abundant microorganism. In the camphor pine forests (CPF) and theropencedrymion (TH), Luteibactor rhizovicina is the largest proportion of microorganisms. Relevant analysis shows that several identified strains from the four forest types, including Bacillus pseudomycoides , Bacillus subtilis , Pseudomonas alcaliphila , Luteibacter rhizovicinus and Pseudomonas alcaliphila can promote the biosynthesis and accumulation of monomeric saponins Rc, Rb1, Rb2, Rb3, Rg2, Rb3, and Rh4. Our research findings emphasize the crucial role of different forest stand types in soil microbial community structure, and explore the accumulation mechanism of ginsenosides from a microbial perspective. In summary, this study provides more theoretical basis for the relationship between different forest types and the bioactive components of medicinal plants. Forest types Ginseng Saponin accumulation Soil microorganism Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Introduction The rhizosphere microorganisms of plants are considered as the "second genome of plants"(Wang et al. 2024 ).The growth, development, yield, secondary metabolites, and disease defense of medicinal plants are crucial. The community structure and diversity of rhizosphere microorganisms are influenced by soil environment, which is related to factors such as forest type. Different forest types have different plant root exudates and accumulated humus, resulting in different soil environments(Man et al. 2023 ).Studying the rhizosphere microorganisms of medicinal plants under different forest types and their impact on the accumulation of effective components in medicinal plants is of great significance. Panax ginseng C.A. Mey. is a perennial perennial herbaceous plant of the Panax genus in the Araliaceae family. It has a history of more than 5000 years of medicinal use in China and is commonly known as "stick hammer"(Potenza et al. 2023 ).It has traditional effects such as restoring meridians, strengthening the spleen and benefiting the lungs, and greatly nourishing vital energy(Jaiswal et al. 2016 ). Plant chemistry studies have shown that the main active ingredient of ginseng is the triterpenoid compound ginsenoside(Kim et al. 2017 ). Modern pharmacological studies have shown that ginsenosides have good anti-tumor, anti-inflammatory, antioxidant, and anti apoptotic effects(Gao et al. 2018 ).With the increasing awareness of health among people, the demand for ginseng is also gradually increasing. The cultivation methods of ginseng mainly include three types: farmland ginseng, garden ginseng, and mountainous forest cultivated ginseng(MFCG). MFCG, also known as cultivated mountain ginseng or seed sea, is divided into forest seed ginseng and forest transplanted ginseng. MFCG is a wild mountain ginseng that artificially grows ginseng seeds or seedlings in the mountains and forests. Compared with the other two cultivation methods of ginseng, the growth environment of ginseng under the forest is closer to that of wild mountain ginseng, and the quality of MFCG for more than ten years can be comparable to that of wild mountain ginseng. The content and type of ginsenosides are important indicators for evaluating ginseng. Although the yield and quality of ginsenosides have been widely studied, the yield of ginsenosides is still very low. Therefore, it is urgent to explore the cumulative impact mechanism of ginsenosides and improve the yield of ginsenosides. The cultivation history of ginseng is long, and China is the country with the largest ginseng planting area in the world(Potenza et al. 2023 ). The Changbai Mountain area in the eastern part of Jilin Province is the main production area of ginseng in China. With the rapid development of the ginseng industry, the planting area of MFCG is also significantly increasing. Ecological factors such as forest type are the main influencing factors on the accumulation of ginsenosides in understory ginseng. Currently, research on the active ingredients of ginseng mainly focuses on pharmacological activity and quality control. However, in terms of microecology, there are currently few literature reports on the mechanism by which forest types affect the accumulation of ginsenosides. Therefore, studying the mechanism of different forest types affecting the accumulation of ginsenosides in MFCG is particularly important for improving the quality of ginseng. The ecosystems under different forest types are also different, and various ecological factors interact and interweave vertically and horizontally, forming a complex network. The ecological networks constructed by different tree species also have significant differences, such as the physical and chemical properties, microbial types and quantities, canopy closure, etc. of the soil under the forest(Bei et al. 2023 ), which leads to forest types becoming one of the important factors affecting the accumulation of ginsenosides. Studying the microbial effects of different forest types on the accumulation of ginsenosides is not only helpful for improving the quality of MFGG, but also crucial for agricultural development and ecological balance. The aim of this study is to reveal the specific differences in soil microbial activity and quantity among four forest types in the Changbai Mountain area: TH, LF, BF, and CPF. Dilution culture and top purification methods were used to isolate and identify soil microorganisms in the rhizosphere of ginseng under different forest types. At the same time, the content of 21 ginsenosides in ginseng was determined by high-performance liquid chromatography (HPLC). Through correlation analysis, the microbial accumulation mechanism of ginsenoside content influenced by forest type was revealed, thereby improving ginsenoside content. Provide theoretical basis for promoting the accumulation of effective ingredients in medicinal materials by rhizosphere microorganisms. Materials and methods S tudy site This study was conducted in the mountainous areas of Jingyu County, Jilin Province, including CPF, BF, LF, and TH(longitude 126 ° 30 '-127 ° 16'N, latitude 42 ° 06' -42 ° 48'E ，fig1.). Jingyu County is located in the hinterland of Changbai Mountain, with a forest coverage rate of up to 84%. It is the main production area of ginseng in Jilin Province and a representative area for studying forest ginseng. This mountainous area belongs to the Changbai Mountain Range. The mountainous area belongs to a continental monsoon climate in the cold temperate zone, with long and cold winters and short and warm and humid summers, with moderate sunlight. Altitude: 475 meters. Annual average temperature: 3.7 ℃, highest temperature is 37 °C, lowest temperature is -48 °C, annual average precipitation is 749.7 mm, and soil depth exceeds 25 centimeters. Table 1 summarizes the quality of the four forest types selected in this article. Table 1. Characteristics of four common planting forest types of understory ginseng in Jingyu County, Jilin Province. Notes:TH，theropencedrymion ；LF，larch forest ；BF，broad-leaved forest；CPF camphor pine forest Sample collection Nine 15-year-old disease-free MFCG from four different forest types were selected. The non-medicinal parts were removed, and the rhizomes were washed, dried, and ground into a fine powder. The ginsenoside content was then analyzed by crushing (0.425 sieve) the powder from each plant. Soils were collected separately from the four forest types mentioned above. Five sample points were selected in the same plot using the 'S-type sampling method'. Soil humus was removed to a depth of 0-20 cm from the surface. The soil samples from the healthy ginseng rhizosphere under different forest types were collected uniformly, placed in sterile ziplock bags, and mixed to ensure representativeness. The samples were then quickly stored in an icebox and transported to the laboratory where they were immediately stored at -80◦C until microbial analysis was required. Determination of ginsenoside content in MFCG samples The content of ginsenosides Rg1, Re, Rf, Rb1, Rg2, Rg3, Rg5, Rc, F1, Rb2, F2, Rh2, Rd, Rb3, compound K, 20 (R)-Rh1, Rk3, Rh4, original ginsenosides, and original ginsenosides under different forest types were determined by HPLC. Weigh 500 mg of ginseng powder for each of the four forest types. Place each sample in a 10 mL centrifuge tube and add 5.0 mL of chromatographic methanol solution. Seal the tubes, weigh them, and extract the samples using ultrasonication (40 Hz, 100 watts power, 45 min). The samples were left overnight, reweighed and sonicated for another 45 min, and the supernatant was transferred to a centrifuge tube with a sterile syringe and centrifuged at 5000 rpm for 15 min, and then filtered through a 0.22 μM filter into a 2.0 mL sample vial for spare use. Weigh 5 mg each of ginsenoside standards Rg1, Re, Rg2, Rg3, Rg5, Rf, F1, F2, Rc, Rd, Rb1, Rb2, Rb3, Rh2, compound K, 20(R)-Rh1, Rk3, Rh4, protopanaxadiol and protopanaxatriol, and add methanol to formulate the mass concentrations of 1.04, 1.02, 0.98 , 1.00, 1.00, 1.02, 1.00, 1.00, 1.00, 1.02, 1.00, 1.00, 1.00, 1.04, 1.00, 1.00, 1.00, 0.98, 1.00, 1.00, 1.00, 1.02 mg/mL, and then filtrated through 0.45 μM filtration membrane, and set aside. The chromatographic conditions refer to the method established in our laboratory for simultaneous determination of 20 types of ginsenosides. The chromatographic column used in the study was BDS C 18 (250 mm × 4.6 mm, 5 μm). The mobile phase was acetonitrile (A) and water (B) with the gradient elution as follows: 0-40 min, 18% A→ 21%; 40-42 min, 21% A→ 26%; 42-46 min, 26% A→ 32%; 46-66 min, 32% A→ 33.8%; 66-71 min, 33.8% A→ 38%; 71-77.7 min, 38% A→ 49.08%; 77.7-78 min, 49.08% A→ 49.1%; 78-82 min, 49.1% A; 82-83 min, 49.1% A→ 50.6%; 83-88 min, 50.6% A→ 59.6%; 88-89.8 min, 59.6% A→ 64.96%; 89.8-92 min, 64.96% A→ 65%; 92-97 min, 65% A; 97-102 min, 65% A→ 85%; 102-109 min, 85% A; 109-111 min, 85% A→ 18%. The velocity of the mobile phase was 1 ml/min. The detection wavelength was set as 203 nm while the temperature of the chromatographic column was 35 °C. Isolation and purification of soil microorganisms in the rhizosphere of ginseng under different forest types The wet weight of ginseng inter-root soil samples under different forest types was accurately weighed 0.5 g, added with 5 mL of sterile water, and incubated at 28 ℃ for 30 min with shaking, and then diluted according to the 10-fold dilution method, respectively, and 100 μL of bacterial suspension with a concentration of 1×10 -7 was taken to coat the plate, and each concentration was repeated three times, and it was placed in the incubator at a constant temperature of 28 ℃ for 1 d-3 d, and then it was reserved for use. The obtained colonies were isolated and purified according to different morphologies, and placed in an incubator at 28 ℃ for inverted culture for 1-3d to observe and record the morphology of the colonies, and when the purification was complete, the colonies were transferred to the corresponding slant for preservation, and named according to their morphological characteristics. Isolation and identification of soil microorganisms in the rhizosphere of ginseng under different forest types Soil bacteria and fungi DNA were extracted using Bacterial Genomic DNA Extraction Kit (Solarbio, Beijing China) and Fungi Genomic DNA Extraction Kit (Solarbio, Beijing China), respectively. The extracted DNA is purified and stored at -20℃ for future use. PCR amplification of bacterial 16s RNA was performed using bacterial universal primers 1492R and 27F. The primers were provided by Bao Bio-engineering Co., Ltd (Dalian, China) and the sequences were 5'-TACGGCTACCTTGTTACGACTT-3' and 5'-AGAGTTTGATCCTGGGCTCAG-3'. The 20 μL PCR reaction system consists of 12.5 μL of 2×Taq MasterMix for Page, 1 μL of upstream primer, 1 μL of downstream primer, 3 μL of DNA template, and 7.5 μL of deionized water. PCR reaction procedure was: pre denaturation at 95 ℃ for 5 min, denaturation at 95 ℃ for 30 s, then annealing at 57℃/54 ℃ for 30 s, and 30 cycles of extension at 72 ℃ for 90 s, and finally extension at 72 ℃ for 10 min. PCR products were detected by 1% agarose gel electrophoresis to observe whether there were specific target bands. The PCR products were then sequenced, and the sequencing service was provided by Sangon Biotech (Shanghai) Co., Ltd (Shanghai, China). The sequences were analyzed by Basic Local Alignment Search Tool (BLAST) in the National Center for Biotechnology Information (NCBI) database to identify the strains with the highest similarity to each other and to determine the biological classification status of each strain. Correlation analysis of ginseng inter-root soil microorganisms with ginsenosides under different forest types The number of each colony of soil under each forest type obtained above and the measured data of ginsenosides of ginseng under each forest type were entered into SPSS 22.0 (Chicago, USA) software for correlation analysis and Pearson Correlation values were obtained, respectively. GraphPad Prism 8.0.2 (San Diego, USA) was used to map the number of ginseng inter-root microorganisms and the content of active compounds under different forest types. Influence of ginseng inter-root microorganisms on the transformation of ginsenosides under different forest types The seven colonies after 24 h of streaking culture on solid potato dextrose agar (PDA) medium were transferred to conical flasks containing 100 mL of liquid PDA medium with a grafting ring for each single colony respectively, and were reciprocally cultured on a shaker at 28 ℃, 140 r/min for 24 h, and the cultures of Bacillus pseudomycoides,Bacillus subtilis , Pseudomonas alcaliphila , Pseudomonas pseudoalcaligenes , Luteibactor rhizovicina , Bacillus cereus , and Bacillus megaterium were aseptically preserved and stored for spare parts. Weigh 0.5 g of ginseng powder through 20 mesh Pharmacopoeia sieve in 36 test tubes, dry heat sterilize at 80 ℃ for 2 h, add 3 mL of sterile water to each test tube separately with pipette gun, and cool to room temperature. Under aseptic conditions, 2mL of each group of bacterial solution was separately pipetted into the above test tubes. The specific information of the groups and inoculated bacterial solution is shown in Table 2. The tubes were then placed in an incubator at 28 ℃ with shaking, and the changes in ginsenoside content were observed after 8 days of incubation. Table 2 Grouping and bacterial liquid information Groups Bacterial solution Control sterile water Bacillus pseudomycoides Bacillus pseudomycoides Bacillus subtilis Bacillus subtilis Pseudomonas alcaliphila Pseudomonas alcaliphila Pseudomonas pseudoalcaligene Pseudomonas pseudoalcaligene Luteibactor rhizovicina Luteibactor rhizovicina Bacillus cereus Bacillus cereus Bacillus megaterium Bacillus megaterium Larch forest Bacillus subtilis 、 Pseudomonas alcaliphila 、 Pseudomonas pseudoalcaligene 、 Luteibactor rhizovicina 、 Bacillus megaterium Camphor pine forest Bacillus subtilis 、 Pseudomonas alcaliphila 、 Pseudomonas pseudoalcaligene 、 Luteibactor rhizovicina 、 Bacillus cereus 、 Bacillus megaterium Broad-leaved forest Bacillus pseudomycoides 、 Bacillus subtilis 、 Pseudomonas alcaliphila 、 Pseudomonas pseudoalcaligene 、 Luteibactor rhizovicina 、 Bacillus megaterium Theropencedrymion Bacillus pseudomycoides 、 Bacillus subtilis 、 Pseudomonas alcaliphila 、 Luteibactor rhizovicina 、 Bacillus megaterium S tatistic analysis The obtained data will be statistically analyzed using SPSS 22.0 (Chicago, USA) software. One-way ANOVA was used to analyze the differences in ginsenoside content among different forest types. Mean values were analyzed using Tukey's test. P < 0.05 was considered to indicate significant differences, and all data were expressed as mean ± standard deviation. Results Effect of different forest types on ginsenoside content in ginseng The ginseng under four forest types all contain 14 types of ginsenoside monomers, as shown in Fig. 1 and Fig. 2 . The content of Rg1 and Rf, as well as the sum of 20 monomeric saponins, were significantly higher in the CPF group than in the other three forest types (P < 0.05). The content of Re, Rb2, and Rb3 in the CPF group was significantly higher than that in the LF group and the BF group (P < 0.05), while the differences between the LH group and the other groups were not significant. The Rb1 content in the CPF group was significantly higher than that in the other three forest types (P < 0.05), while Rb1 content was higher (P < 0.05) in the TH and BF groups than in the LF group. The Rg2 content in the LF group was significantly higher than that in the other three forest types (P < 0.05). The Rd content in TH group and the CPF group was significantly higher than the other two groups (P < 0.05). The Rh4 content in the LF group was significantly higher than that in the TH group (P < 0.05). The content of ginsenoside Rg5 in the CPF group and the BF group was significantly higher than the other two groups (P < 0.05). Under the CPF forest type, the saponin content in the ginseng under the forest is the highest. Isolation, purification, and morphological observation of soil microorganisms in the rhizosphere of ginseng under different forest types As shown in Fig. 3 and Fig. 4 , separate and purify soil microorganisms to obtain 7 bacterial strains. JQ.GSRS-1: The colonies are white, rod-shaped, with round ends and arranged in a chain like manner. JQ.GSRS-2: The colony is dirty white, elliptical to columnar in shape, with a rough and opaque surface that forms wrinkles. JQ.GSRS-3: The colony is white, rod-shaped, and has flagella at the end, which can move. JQ.GSRS-4: The colony is green with flagella, round in shape, with neat edges and a smooth surface. JQ.GSRS-5: The colony is yellow in color, round in shape, raised, and sticky, making it easy to pick up. JQ.GSRS-6: The colony is in the shape of ground glass, the spores are round, the surface is rough, flat, and irregular, forming a snowflake like shape. JQ.GSRS-7: The colony is milky white, rod-shaped, and has a round end. The number of bacteria in the rhizosphere soil of ginseng under different forest types As shown in Fig. 6 the statistical results of the number of microorganisms in the rhizosphere soil of different forest types of ginsengs showed that a total of 7 bacteria were isolated and identified in the rhizosphere soil of four different forest types of MFCG. Among them, Bacillus pseudomycoides and Bacillus cereus were not isolated and identified in the MFCG soil of the LF type; Pseudomonas aeruginosa was not isolated and identified in the MFCG soil of CPF type; Pseudomonas pseudoalcaligenes and Bacillus cereus were not isolated and identified in the MFCG soil of the TH type. Bacillus subtilis , Pseudomonas alcaliphila , Luteibacter rhizovicinus and Bacillus megaterium were detected in all four types of forest MFCG soils. Bacillus pseudomycoides was isolated and identified in BF and TH, with a quantity of 0.33 in the BF(×10 6 cfu/g), the number of Bacillus pseudomycoides in the TH group is 0.35༈×10 6 cfu/g), there was no significant difference in the number of Bacillus pseudomycoides between the two forest types. The number of Bacillus subtilis in BF is 5.01(×10 6 cfu/g), the number of Bacillus subtilis in the LF is 4.67༈×10 6 cfu/g), the number of Bacillus subtilis in the CPF is 5.59༈×10 6 cfu/g), the number of Bacillus subtilis in TH is 6.34༈×10 6 cfu/g), there was no significant difference in the number of Bacillus subtilis among the four forest types. The number of Pseudomonas alcaligenes in BF is 3.57(×10 6 cfu/g), the number of Pseudomonas alcaligenes in the LF is 0.69༈×10 6 cfu/g), the number of Pseudomonas alcaligenes in the CPF is 2.35༈×10 6 cfu/g), the number of Pseudomonas alcaligenes in the TH is 0.68༈×10 6 cfu/g), the content of Pseudomonas alcaligenes is higher in BF than in the other three forest types. The number of Pseudomonas alcaligenes is similar and smaller in LF and TH than in CPF. Pseudomonas pseudoalcaligenes was detected in three forest types: BF, LF, and CPF. The number of Pseudomonas pseudoalcaligenes in the LF group was 9.57(×10 6 cfu/g), higher than the BF group and the CPF group, with a content of 2.03༈×10 6 cfu/g) Pseudomonas pseudoalcaligenes in the BF group, the content of Pseudomonas pseudoalcaligenes in the CPF group is 3.47༈×10 6 cfu/g). The number of Luteibactor rhizovicina in the CPF is 38.13(×10 6 cfu/g, significantly higher than the other three forest types, with a population of 17.69 Luteibactor rhizovicina in BF༈×10 6 cfu/g), the number of Luteibactor rhizovicina in the LF is 12.03༈×10 6 cfu/g), the number of Luteibactor rhizovicina in the TH is 17.35༈× 10 6 cfu/g). Wax like Bacillus cereus was detected in the CPF group, with a quantity of 4.3333(×10 6 cfu/g), no Bacillus cereus was detected in the other three forest types. The number of Bacillus megaterium in the LF is 23.47(×10 6 cfu/g), significantly higher than the other three forest type groups. The number of Bacillus megaterium in the BF is 18.05 (× 10 6 cfu/g).The number of Bacillus megaterium in the CPF is 12.02 (× 10 6 cfu/g), and in the TH, the number of Bacillus megaterium is 8.37 (× 10 6 cfu/g). The effect of ginseng rhizosphere microorganisms on the transformation of ginsenosides under different forest types To verify the effect of different fungal species in the rhizosphere of ginseng under different forest types on the content of ginsenosides, a control group was set up based on the 7 identified bacteria in the bacteria detected under different forest types, Set up control groups separately, Bacillus pseudomycoides group; Bacillus subtilis group; Pseudomonas alcaliphila group; Pseudomonas pseudoalcaligenes group; Luteibactor rhizovicina group; Bacillus cereus group; Bacillus megaterium group.Set up LF groups according to different forest types(mixed of Bacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus megaterium ),CPF group (mixed with Bacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus cereus、Bacillus megaterium );BF group (mixed with Bacillus pseudomycoides、Bacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus megaterium ); TH group ( mixed with Bacillus pseudomycoides、Bacillus subtilis、Pseudomonas alcaliphila、Luteibactor rhizovicina、Bacillus megaterium ). The results of ginsenoside detection in each group are shown in Fig. 7 and the 7 isolated and identified bacteria have different promoting effects on the monomer saponins. The group of Bacillus pseudomycoides had a significant promoting effect on the monomer saponin Rc (P < 0.05); The Bacillus subtilis group had a good promoting effect on the monomeric saponins Rg2, Rb2, and Rd (P < 0.05); The promotion effect of the Pseudomonas alcaliphila group on the monomer saponin Rb3, diol type saponin, and saponin addition value is more significant (P < 0.05); The group of Pseudomonas pseudoalcaligenes showed a significant promoting effect on the monomer saponin Rh2 (P < 0.05); The group of Luteibactor rhizovicina had a significant promoting effect on the monomer saponins Rg2, Rb3, F2, Rh4, diol type saponins, triol type saponins, and saponin addition values (P < 0.05); The promotion effect of Bacillus cereus group on monomeric saponins Rc and Rb2 was more significant (P < 0.05); The Bacillus megaterium group also has a good promoting effect on monomeric saponins Rc and Rb2. The mixed microbial colonies in ginseng soil under LF and TH types have a good promoting effect on the monomeric saponins Rc, Rb3, and diol type saponins (P < 0.05); The CPF group had a significant promoting effect on the monomer saponin Rh4 (P < 0.05); The BF group has a significant promoting effect on the monomer saponin Rb3 and diol type saponins (P < 0.05). The correlation between the accumulation of ginsenosides in different forest types and the rhizosphere soil microorganisms of MFCG The correlation analysis results (fig.8) indicate that there is varying degrees of correlation between the content of different monomeric saponins and the number of microorganisms, with the highest positive correlation coefficient between monomeric saponin Rg1 and Bacillus subtilis (r=0.5987). The negative correlation coefficient between Rg1 and Pseudomonas alcaliphila was the highest (r=-0.7603), and the negative correlation coefficients of other soil microorganisms were ranked as follows: Bacillus pseudomycoides strain > Bacillus cereus > Bacillus megaterium > Pseudomonas pseudoalcaligenes. The positive correlation coefficient between monomer saponin Re and Bacillus subtilis is the highest (r=0.7820). The negative correlation coefficient between Re and Pseudomonas pseudoalcaligenes was the highest (r=-0.8717), and the negative correlation coefficients of other soil microorganisms were ranked as follows: Bacillus megaterium > Pseudomonas pseudoalcaligenes > Luteibactor rhizovicina > Pseudomonas alcaliphila. The positive correlation coefficient between the monomer saponin Rf and Bacillus subtilis was the highest (r=0.7820), and the negative correlation coefficient between Rf and Bacillus megaterium was the highest (r=-0.8530). The negative correlation coefficient of other soil microorganisms was ranked as follows: Bacillus pseudomycoides > Pseudomonas pseudoalcaligenes > Pseudomonas alcaliphila > Bacillus cereus. The single saponin Rb1 has the highest positive correlation coefficient with Bacillus subtilis (r=0.9510). The negative correlation coefficient between Rb1 and Bacillus megaterium was the highest (r=-0.9560), and the negative correlation coefficients of other soil microorganisms were ranked as follows: Pseudomonas pseudoalcaligenes > Bacillus pseudomycoides > Bacillus cereus > Pseudomonas alcaligenes. The single saponin Rg2 has the highest positive correlation coefficient with Luteibacter rhizovicinus (r=0.9736), while the other soil microorganisms have the following positive correlation coefficients: Pseudomonas pseudoalcaligenes > Bacillus subtilis > Bacillus megaterium > Bacillus subtilis > Bacillus pseudomycoides . The negative correlation coefficient between Rg2 and Pseudomonas alcaliphila is the highest (r=-0.5280). The positive correlation coefficient between the monomer saponin Rc and Bacillus megaterium was the highest (r=0.9630), and the positive correlation coefficient of other soil microorganisms was ranked as follows: Bacillus pseudomycoides > Bacillus subtilis . The negative correlation coefficient between Rc and Luteibactor rhizovicina is the highest (r=-0.6602), and the negative correlation coefficient of other soil microorganisms is ranked as follows: Pseudomonas alcaliphila > Pseudomonas pseudoalcaligenes > Bacillus megaterium . The positive correlation coefficient between monomer saponin Rb2 and Bacillus subtilis is the highest (r=0.9450). The negative correlation coefficient between Rb2 and Pseudomonas pseudoalcaligenes was the highest (r=-0.7286), and the negative correlation coefficients of other soil microorganisms were ranked as follows: Bacillus megaterium > Luteibactor rhizovicina > Bacillus pseudomycoides > Pseudomonas alcaliphila. The positive correlation coefficient between monomeric saponin Rb3 and Pseudomonas alcaliphila was the highest (r=0.9710), and the positive correlation coefficient of other soil microorganisms was ranked as follows: Luteibacter rhizovicinus > Bacillus subtilis > Bacillus cereus . The negative correlation coefficient between Rb3 and Pseudomonas pseudoalcaligenes was the highest (r=-0.7374), and the negative correlation coefficient of other soil microorganisms was ranked as follows: Bacillus megaterium > Bacillus pseudomycoides . The positive correlation coefficient between the monomer saponin Rd and Bacillus cereus was the highest (r=0.5722), and the positive correlation coefficient of other soil microorganisms was ranked as follows: Bacillus subtilis > Bacillus pseudomycoides > Pseudomonas alcaliphila . Rd has the highest negative correlation coefficient with Luteibacter rhizovicinus （r=-0.6132）,and the negative correlation coefficient of other soil microorganisms was ranked as follows: Pseudomonas pseudoalcaligenes > Bacillus megaterium. The positive correlation coefficient between Rh4 and Luteibacter rhizovicinus was the largest (r=0.9590), and the positive correlation coefficients of other soil microorganisms were in the order of Pseudomonas pseudoalcaligenes > Bacillus megaterium > Bacillus pseudomycoides . Rh4 had the largest negative correlation coefficient with Pseudomonas alcaliphila (r=-0.7875), and the other soil microorganisms were in the order of Bacillus subtilis > Bacillus cereus . The positive correlation coefficient between Compound K and Pseudomonas alcaliphila was the largest (r=0.7413), and the positive correlation coefficients of other soil microorganisms were in the order of Bacillus megaterium > Bacillus pseudomycoides . Compound K had the largest negative correlation coefficient with Pseudomonas alcaliphila (r=-0.7863), and the other soil microorganisms were in the order of Bacillus subtilis > Luteibacter rhizovicinus > Bacillus cereus . The positive correlation coefficient between Rg5 and Luteibacter rhizovicinus was the highest (r=0.7699). The negative correlation coefficient between Rg5 and Bacillus pseudomycoides was the largest (r=-0.9730), and the negative correlation coefficients of other soil microorganisms were in the order of Bacillus megaterium > Bacillus cereus > Pseudomonas pseudoalcaligenes > Pseudomonas alcaliphila . The positive correlation coefficient between Rh2 and Pseudomonas pseudoalcaligenes was the largest (r=0.8818), and the positive correlation coefficients of other soil microorganisms were in the order of Bacillus megaterium > Bacillus pseudomycoides . Rh2 had the largest negative correlation coefficient with Pseudomonas alcaliphila (r=-0.7088), and the other soil microorganisms were in the order of Bacillus subtilis > Bacillus cereus > Luteibacter rhizovicinus . The positive correlation coefficient between propanaxanediol and Bacillus subtilis was the largest (r=0.8496). The negative correlation coefficient between propanaxanediol and Bacillus megaterium was the largest (r=-0.8784), and the negative correlation coefficients of other soil microorganisms were in the order of Bacillus pseudomycoides > Pseudomonas pseudoalcaligenes > Pseudomonas alcaliphila > Bacillus cereus . The positive correlation coefficient between diol type and Pseudomonas alcaliphila was the largest (r=0.9620). The negative correlation coefficient between diol type and Bacillus megaterium was the largest (r=-0.8304), and the negative correlation coefficients of other soil microorganisms were in the order of Pseudomonas pseudoalcaligenes > Bacillus pseudomycoides > Luteibacter rhizovicinus > Bacillus cereus . The positive correlation coefficient between triol type and Luteibacter rhizovicinus was the largest (r=0.9339), and the positive correlation coefficient of other soil microorganisms was in the order of Pseudomonas pseudoalcaligenes > Bacillus subtilis . The negative correlation coefficient between triol type and Bacillus cereus was the largest (r=-0.6669), and the negative correlation coefficient of other soil microorganisms was in the order of Pseudomonas alcaliphila > Bacillus pseudomycoides > Bacillus megaterium . The positive correlation coefficient between the addition value of monomonins and Luteibacter rhizovicinus was the largest (r=0.9633). The negative correlation coefficient between the sum of monomonins and Pseudomonas pseudoalcaligenes was the largest (r=-0.6015), and the other negative correlation coefficients of soil microorganisms were in the order of Bacillus pseudomycoides > Pseudomonas alcaliphila > Bacillus cereus . Discussion Ginseng ( Panax ginseng C.A. Meyer), as a representative rare traditional medicinal plant, has been eaten in China for more than 5000 years(Potenza et al. 2023). At present, studies have proved that ginseng has a good therapeutic effect on fatigue, anorexia, shortness of breath, palpitations, insomnia and diabetes(Gao et al. 2018). Due to its remarkable pharmacological properties, ginseng has been widely recognized in Asia and attracted a large number of consumer groups, but excessive mining has also led to the rapid extinction of wild ginseng resources in just a few decades(L 2015). In order to meet the increasing market demand, people began to widely plant ginseng under the forest to replace wild ginseng. MFCG is a semi wild ginseng cultivated by artificially sowing ginseng seedlings or seeds under natural or artificial forests, simulating the growth process of wild ginseng. Its appearance and composition are similar to some characteristics of wild ginseng. China has been trying to cultivate ginseng under the forest in the Northeast region since the 1980s(F 2019). In recent years, in accordance with the requirements of China's ecological civilization construction policy regarding the development of understory economy, promoting understory ginseng cultivation has become the main trend in the development of ginseng cultivation in Northeast China. By relying on a series of industrial support policies from the government for the cultivation of ginseng under forests, the scale of ginseng cultivation in China has been further expanded(Liu et al. 2016). The pharmacological effects of ginseng are closely related to its multiple active ingredients in the body, including ginsenosides, polysaccharides, peptides, plant sterols, etc. Among them, ginsenosides are the main pharmacological active ingredients of ginseng. Ginsenoside belongs to the triterpenoid class of compounds, which are a type of terpenoid compounds composed of 30 carbon atoms in the basic nucleus, and are polymerized from 6 isoprene units. There are currently over 100 known types of ginsenosides, which can be divided into tetracyclic triterpenes and pentacyclic triterpenes based on their different mother ring structures. Ginsenoside has high medicinal activity and its pharmacological effects have been widely studied in recent years. The research results indicate that it has the effects of enhancing physical strength, improving immunity, and prolonging life. At the same time, it can also resist a variety of adverse reactions, including depression, diabetes, fatigue, aging, inflammation, tumors, lung problems, indigestion, vomiting, tension, stress, ulcers and other adverse reactions(Wang et al. 2009). Based on the rich pharmacological activity of ginsenosides, how to improve the content of ginsenosides in ginseng medicinal materials has always been a hot research topic for scholars. However, due to the complex transformation and accumulation mechanism of ginsenosides in ginseng, there is still no good solution to this problem. The main factors affecting the accumulation of ginsenosides in MFCG are ecological factors such as forest type(Wang et al. 2021). However, the specific ways in which different forest types regulate the content of ginsenosides in MFCG have not been thoroughly studied. In recent years, an increasing number of studies have shown that soil rhizosphere microorganisms have a significant impact on the increase of effective ingredient content in medicinal plants(Wang et al. 2021). However, there has been no in-depth research on whether the rhizosphere soil microorganisms of Panax ginseng in different forest types are involved in the transformation and accumulation of ginsenosides in Panax ginseng. In this study, rhizosphere soil and MFCG samples were collected from four different forest types (CPF, BF, LF, and TH) in the mountainous area of Jingyu County, Jilin Province, China. Subsequently, the content of 20 ginsenosides in the MFCG samples under the forest was analyzed using high-performance liquid chromatography, and the microorganisms in the ginseng rhizosphere soil were isolated, purified, and identified. Based on the experimental results, we conducted a series of correlation analyses, including: correlation analysis between forest type and rhizosphere soil microorganisms, correlation analysis between rhizosphere soil microorganisms and ginsenoside transformation in ginseng under the forest, correlation analysis between rhizosphere soil microorganisms and ginsenoside accumulation in ginseng under the forest, etc. Effects of different forest types on MFCG quality Therefore, we selected several representative forest types planted under the forest type of understory ginseng as the research object, and determined the content of 20 important ginseng monomer saponins (Rg1, Re, Rf, Rb1, Rg2, Rc, Rh1, Rb2, Rb3, F1, Rd, Rk3, F2, Rh4, Rg3, There were 14 ginsenosides (Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, Rd, Rh4, Compound K, Rg5, Rh2, Progindadiol) in ginseng under the four forest types, but there were some differences in the content of monomeric saponins and the addition value of 14 saponins. Among them, the sum value of 14 monomeric saponins in Pinus sylvestris var. mongolica was the highest, and the quality was the best, so we believe that CPF was the most suitable for planting ginseng. The sum value of the 14 monomeric saponins of ginseng under the BF was the lowest.However, there was no change in the monomeric saponin species，This may be related to G × E (genotype × environment) interactions.For example, the activity of dogwood, basil, ashwagandha and notoginseng(Liu et al. 2018; Xu et al. 2023; Kumar et al. 2023).In addition, widely distributed species in order to adapt to different environments lead to the creation of stable genetic variation, resulting in multiple genotypes of the same species. It is possible that the composition and chemical content of these genotypes will be different, for example, trans-anethole, the main ingredient in fennel, is higher in both genotypes and there is no difference between the other genotypes(Yaldiz and Camlica 2019).We hypothesized that environmental variability may be one of the factors contributing to the significant differences in the content of ginsenosides, the main active ingredient of ginseng. This is also confirmed by other people's reports(Zhu et al. 2022).Indicates that differences in environmental conditions, as well as soil properties, may lead to substantial differences in the quality of medicinal plant material. Effects of different forest types on soil microorganisms in the rhizosphere of ginseng Microorganisms in the soil play the role of decomposers in the forest ecosystem, and participate in the process of information transmission and energy flow in the forest(Li et al. 2021; Kunito et al. 2012).A total of 7 bacteria ( Bacillus pseudomycoides 、 Pseudospora subtilis 、 Pseudomonas alcaliphila 、 Pseudomonas pseudoalcaligenes 、 Luteibacter rhizovicinus 、 Bacillus cereus and Bacillus megaterium ) were isolated and identified in the rhizosphere soil of ginseng in four different forest types, among which the number of microorganisms was the highest in the BF soil, the highest number of Luteibactor rhizovicina in the pine of CPF, and there was no significant difference in the number of colonies in the LF and the TH. The results showed that there was a strong correlation between forest type and soil microorganisms, which was consistent with the results of previous studies(Liu et al. 2023).Soil microorganisms are closely related to the synthesis of ginsenosides, and Wang et al. studied that Aspergillus niger fungi can promote the production of ginsenosides(Wang et al. 2019).There is also Penicillium YJM-2013 that promotes the accumulation of ginsenosides by enhancing the production of signaling molecules, activating the expression of transcription factors and functional genes(Wang et al. 2020).Four bacteria ( Pseudospora subtilis , Pseudomonas alcaliphila , Luteibacter rhizovicinus Bacillus megamegasporus ) were detected in four different forest types. Bacillus cereus was detected only in the rhizosphere soil of ginseng under CPF.We found that therefore, we believe that forest type has a greater impact on the species and number of soil bacteria in the rhizosphere of ginseng. Therefore, we believe that the number and species of rhizosphere microorganisms in the soil of ginseng under the forest type were affected by the forest type, and Bacillus cereus promoted the accumulation of ginsenosides. Effects of rhizosphere soil microorganisms on medicinal plants Correlation coefficient between ginsenoside accumulation and soil microorganisms in the rhizosphere of ginseng according to different forest types (fig. 8),It was found that the content of ginsenosides was more closely related to the types and quantities of soil bacteria, indicating that they played an important role in the formation of ginseng quality. Soil rooting bacteria affect the colonization of plant roots by growth-promoting bacteria and stimulate plant growth and development. Recent studies have proved that rhizosphere bacteria and secondary metabolites not only affect plant growth and resistance to stress stress, but also affect the absorption of nutrients and the accumulation of active ingredients in medicinal plants(Yang et al. 2009; Dhungana et al. 2023; Shang et al. 2023).For example, preliminary experiments in our laboratory have confirmed that Bacillus cereus have a disease prevention and growth promotion effect on ginseng(Y 2023).Relevant representative studies have shown that moss and Bacillus subtilis can significantly increase the content and yield of artemisinin(Awasthi et al. 2011).Trichosa can increase the production of ginsenosides through biological induction(Xu et al. 2021). Therefore, we speculated that ginseng under different forest types may selectively recruit specific microorganisms to metabolize certain substances, thereby affecting ginsenoside content.Our experimental results showed that seven different bacteria were isolated from the four forest types.The enrichment of seven bacteria, including Bacillus pseudomycoides, Bacillus subtilis,Pseudomonas alcaliphila,Pseudomonas pseudoalcaligenes, Luteibacter rhizovicinus Bacillus cereus, and Bacillus megaterium, may promote the biosynthesis and accumulation of ginsenosides, the triterpenoid glycoside compounds of ginseng.The mechanism of the effect of different forest types on microbial-mediated ginsenoside accumulation was clarified. Declarations Author contributions Conceptualization;Yugang Gao. Funding acquisition;Yugang Gao. Investigation; Fengyu Pang, Xiaojia Ruan, Qun Liu. Methodology;Yugang Gao, Yan Zhao, Fengyu Pang, Qun Liu. Supervision; Yugang Gao,Yan Zhao. Visualization; Fengyu Pang. Roles/Writing - original draft; Fengyu Pang, Xiaojia Ruan.writing - review & editing; Fengyu Pang, Xiaojia Ruan, Yan Zhao, Yugang Gao,Qun Liu. Funding We thank funding supports for current research from National Key Research and Development Programme (Grant No. 2022YFF1300503) and Science and Technology Development Program of Jilin Province (Grant No. 20220401110YY). Data availability Data will be made available on request. Conflict of interest The authors declare no conflict of interest. References Awasthi A, Bharti N, Nair P, Singh R, Shukla AK, Gupta MM, Darokar MP, Kalra A (2011) Synergistic effect of Glomus mosseae and nitrogen fixing Bacillus subtilis strain Daz26 on artemisinin content in Artemisia annua L. Applied Soil Ecology 49:125-130. doi:https://doi.org/10.1016/j.apsoil.2011.06.005 Bei Q, Yang T, Ren C, Guan E, Dai Y, Shu D, He W, Tian H, Wei G (2023) Soil pH determines arsenic-related functional gene and bacterial diversity in natural forests on the Taibai Mountain. Environmental Research 220:115181. doi:https://doi.org/10.1016/j.envres.2022.115181 Dhungana I, Kantar MB, Nguyen NH (2023) Root exudate composition from different plant species influences the growth of rhizosphere bacteria. Rhizosphere 25:100645. doi:https://doi.org/10.1016/j.rhisph.2022.100645 F H (2019) Transeriptional analysis of terpenoids metabolism in developing Panax ginseng root Beijing Forestry University Gao Y, Liu J, Ji Q, Zhao Y, Zang P, He Z, Zhu H, Zhang L (2018) Anti-tumor activity and related mechanism study of Bacillus Polymyxa transformed Panax ginseng C. A. Mey. Process Biochemistry 72:198-208. doi:https://doi.org/10.1016/j.procbio.2018.06.013 Jaiswal Y, Liang Z, Zhao Z (2016) Botanical drugs in Ayurveda and Traditional Chinese Medicine. Journal of Ethnopharmacology 194:245-259. doi:https://doi.org/10.1016/j.jep.2016.06.052 Kim JH, Yi Y-S, Kim M-Y, Cho JY (2017) Role of ginsenosides, the main active components of Panax ginseng, in inflammatory responses and diseases. Journal of Ginseng Research 41 (4):435-443. doi:https://doi.org/10.1016/j.jgr.2016.08.004 Kumar A, Venugopal S, Jnanesha AC, Lal RK (2023) Agricultural-based challenges, genetic enhancement, and obstacles to an industrially important medicinal plant, ashwagandha (Withania somnifera (L.) Dunal): A review. Ecological Genetics and Genomics 28:100183. doi:https://doi.org/10.1016/j.egg.2023.100183 Kunito T, Tobitani T, Moro H, Toda H (2012) Phosphorus limitation in microorganisms leads to high phosphomonoesterase activity in acid forest soils. Pedobiologia 55 (5):263-270. doi:https://doi.org/10.1016/j.pedobi.2012.05.002 L X (2015) Research on Prediction and Evaluation of Light Environment of Cultivated Ginseng under Forest. Jilin University Li J, Zhu T, Singh BK, Pendall E, Li B, Fang C, Nie M (2021) Key microorganisms mediate soil carbon-climate feedbacks in forest ecosystems. Science Bulletin 66 (19):2036-2044. doi:https://doi.org/10.1016/j.scib.2021.03.008 Liu C, Zheng C, Wang L, Zhang J, Wang Q, Shao S, Qin H, Xu Q, Liang C, Chen J (2023) Moso bamboo invasion changes the assembly process and interactive relationship of soil microbial communities in a subtropical broadleaf forest. Forest Ecology and Management 536:120901. doi:https://doi.org/10.1016/j.foreco.2023.120901 Liu W-w, Liu M-c, Li W-h, Zeng F-s, Qu Y (2016) Influence of ginseng cultivation under larch plantations on plant diversity and soil properties in Liaoning Province, Northeast China. Journal of Mountain Science 13 (9):1598-1608. doi:10.1007/s11629-015-3753-x Liu Y-R, Delgado-Baquerizo M, Wang J-T, Hu H-W, Yang Z, He J-Z (2018) New insights into the role of microbial community composition in driving soil respiration rates. Soil Biology and Biochemistry 118:35-41. doi:https://doi.org/10.1016/j.soilbio.2017.12.003 Man H, Dong X, Li M, Zheng Z, Wang C, Zang S (2023) Spatial distribution and influencing factors of humus layer thickness of forest land in permafrost region of Northeast China. CATENA 224:106979. doi:https://doi.org/10.1016/j.catena.2023.106979 Potenza MA, Montagnani M, Santacroce L, Charitos IA, Bottalico L (2023) Ancient herbal therapy: A brief history of Panax ginseng. Journal of Ginseng Research 47 (3):359-365. doi:https://doi.org/10.1016/j.jgr.2022.03.004 Shang X-c, Zhang M, Zhang Y, Hou X, Yang L (2023) Waste seaweed compost and rhizosphere bacteria Pseudomonas koreensis promote tomato seedlings growth by benefiting properties, enzyme activities and rhizosphere bacterial community in coastal saline soil of Yellow River Delta, China. Waste Management 172:33-42. doi:https://doi.org/10.1016/j.wasman.2023.09.003 Wang B, Chen C, Xiao Y-M, Chen K-Y, Wang J, Zhao S, Liu N, Li J-N, Zhou G-Y (2024) Trophic relationships between protists and bacteria and fungi drive the biogeography of rhizosphere soil microbial community and impact plant physiological and ecological functions. Microbiological Research 280:127603. doi:https://doi.org/10.1016/j.micres.2024.127603 Wang CQ, Yi LW, Zhao L, Zhou YZ, Guo F, Huo YS, Zhao DQ, Xu F, Wang X, Cai SQ (2021) 177 Saponins, Including 11 New Compounds in Wild Ginseng Tentatively Identified via HPLC-IT-TOF-MS(n), and Differences among Wild Ginseng, Ginseng under Forest, and Cultivated Ginseng. Molecules 26 (11). doi:10.3390/molecules26113371 Wang S-h, Liang W-x, Lu J, Yao L, Wang J, Gao W-y (2020) Penicillium sp. YJM-2013 induces ginsenosides biosynthesis in Panax ginseng adventitious roots by inducing plant resistance responses. Chinese Herbal Medicines 12 (3):257-264. doi:https://doi.org/10.1016/j.chmed.2020.02.003 Wang S, Wang J, Liang W, Yao L, Gao W (2019) Promotion of ginsenosides production in a co-cultivation system of Panax ginseng adventitious roots and immobilized Aspergillus niger. Industrial Crops and Products 140:111564. doi:https://doi.org/10.1016/j.indcrop.2019.111564 Wang Y-Z, Chen J, Chu S-F, Wang Y-S, Wang X-Y, Chen N-H, Zhang J-T (2009) Improvement of Memory in Mice and Increase of Hippocampal Excitability in Rats by Ginsenoside Rg1’s Metabolites Ginsenoside Rh1 and Protopanaxatriol. Journal of Pharmacological Sciences 109 (4):504-510. doi:https://doi.org/10.1254/jphs.08060FP Xu X-d, Liang W-x, Yao L, Paek K-Y, Wang J, Gao W-y (2021) Production of ginsenoside by Chaetomium sp. and its effect on enhancing the contents of ginsenosides in Panax ginseng adventitious roots. Biochemical Engineering Journal 174:108100. doi:https://doi.org/10.1016/j.bej.2021.108100 Xu Y, Zhu M, Feng Y, Xu H (2023) Panax notoginseng-microbiota interactions: From plant cultivation to medicinal application. Phytomedicine 119:154978. doi:https://doi.org/10.1016/j.phymed.2023.154978 Y T (2023) The effect of symbiotic bacteria and trace elements on the degradation of pesticide residues by Bacillus polymyxoides. Jilin Agricultural University Yaldiz G, Camlica M (2019) Variation in the fruit phytochemical and mineral composition, and phenolic content and antioxidant activity of the fruit extracts of different fennel (Foeniculum vulgare L.) genotypes. Industrial Crops and Products 142:111852. doi:https://doi.org/10.1016/j.indcrop.2019.111852 Yang J, Kloepper JW, Ryu C-M (2009) Rhizosphere bacteria help plants tolerate abiotic stress. Trends in Plant Science 14 (1):1-4. doi:https://doi.org/10.1016/j.tplants.2008.10.004 Zhu L, Xu L, Huang Y, Xie C, Dou D, Xu J (2022) Correlations between ecological factors and the chemical compositions of mountainous forest cultivated ginseng. Journal of Food Composition and Analysis 114:104867. doi:https://doi.org/10.1016/j.jfca.2022.104867 Supplementary Files AppendixA.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-4487770","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":308956855,"identity":"f78e9935-fb10-4398-bfe0-16ede0b7d885","order_by":0,"name":"Fengyu Pang","email":"","orcid":"","institution":"Jilin Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Fengyu","middleName":"","lastName":"Pang","suffix":""},{"id":308956856,"identity":"137ddfc1-4540-47a4-a19d-f5c94a5f7669","order_by":1,"name":"Xiaojia Ruan","email":"","orcid":"","institution":"Jilin Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Xiaojia","middleName":"","lastName":"Ruan","suffix":""},{"id":308956857,"identity":"4a341cee-88e7-4f4b-a7e1-b753e90c91b3","order_by":2,"name":"Yugang Gao","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBElEQVRIiWNgGAWjYHAD5gMHEipsePj5G4jWwpb44MOZNBnJGQeI1sJjbDiz5bCNQUMCfnXy7oePSfP8ssuTd2Awk+ZtOM9jwHCA8cPHHNxaDM+kpUnz9iUXGx5gADJ23OYxZ25glpy5DY+Whhyg4T3MiRsbGI5J8565zWPZcICNmReflv43IC31QC2MbdK8bed4DA4k4NciLwG0hefH4cT5DMzMhjPbDhDWYiDxLNlybsPxxA0MbIzAQE7mkZxxsBmvX+T7kw/eePOnOnF+A/8HYFTa2fPzNx/88BGfLQcYWCQY24CM+w9gYowNuNWDbGlgYP7A8AfMGAWjYBSMglGAHQAA2+NWw8fhaPoAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0001-5510-5895","institution":"Jilin Agricultural University","correspondingAuthor":true,"prefix":"","firstName":"Yugang","middleName":"","lastName":"Gao","suffix":""},{"id":308956858,"identity":"7bc5f7b2-1680-4ba4-9214-883601688c25","order_by":3,"name":"Yan Zhao","email":"","orcid":"","institution":"Jilin Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Yan","middleName":"","lastName":"Zhao","suffix":""},{"id":308956859,"identity":"6783b86f-027e-4524-b9e0-f1abd545b63f","order_by":4,"name":"Qun Liu","email":"","orcid":"","institution":"Institute of Botany Jiangsu Province and Chinese Academy of Sciences","correspondingAuthor":false,"prefix":"","firstName":"Qun","middleName":"","lastName":"Liu","suffix":""}],"badges":[],"createdAt":"2024-05-28 03:12:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4487770/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4487770/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":58260378,"identity":"0b7a6f62-2a1d-4f17-bfbb-d524ad03e708","added_by":"auto","created_at":"2024-06-13 06:09:01","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":396951,"visible":true,"origin":"","legend":"\u003cp\u003eHPLC chromatogram of the effect of different forest types on the content of MFCG. A. TH; B. LF; C. BF; D. CPF.1-20 is sequentially composed of ginsenosides Rg1, Re, Rf, Rb1, Rg2, Rc, Rh1, Rb2, Rb3, F1, Rd, Rk3, F2, Rh4, Rg3, Original ginseng triol, Compound K, Rg5, Rh2, Protopanaxadiol\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/76489975acfafc8f7e13900a.png"},{"id":58260380,"identity":"e0e595c5-dfbd-42b4-bedf-48f6bf0ab945","added_by":"auto","created_at":"2024-06-13 06:09:02","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":13098,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Different Forest Types on Ginsenoside Content (Unit:%)\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/07c5d6514b677b7ae5927da5.png"},{"id":58260387,"identity":"3b183f81-be40-4ca8-9d9d-96aa56afb5c9","added_by":"auto","created_at":"2024-06-13 06:09:02","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":665415,"visible":true,"origin":"","legend":"\u003cp\u003eA-D represents soil bacteria initially isolated from the rhizosphere soil of ginseng in coniferous and broad-leaved mixed forests, broad-leaved forests, camphor pine forests, and deciduous pine forests.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/4b836757241e3353698f10b0.png"},{"id":58260383,"identity":"e11dfae9-1a82-40b3-9244-2f0737469693","added_by":"auto","created_at":"2024-06-13 06:09:02","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":392275,"visible":true,"origin":"","legend":"\u003cp\u003eMorphological observation on soil microbial separation\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/929eac405dc561722dc88476.png"},{"id":58260865,"identity":"41885893-4e9b-4ac9-9a0f-865157b887ec","added_by":"auto","created_at":"2024-06-13 06:17:02","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":825202,"visible":true,"origin":"","legend":"\u003cp\u003ePathogens homology alignment \u003cem\u003e\u0026nbsp;Pseudomonas alcaliphila strain KR1 16S、KR1 16S； Pseudomonas alcaliphilastrain BC2-110 16S、BC2-110 16S； Pseudomonas alcaliphila strain L59 16S、 L59 16S；Bacillus subtilis strain 261ZG6 16S、261ZG6 16S；Bacillus subtilis strain ZY05 16S、 ZY05 16S；Bacillus subtilis strain Baws1 16S、Baws1 16S； Pseudomonas alcaliphila strain L8 16S、L8 16S； Pseudomonas alcaliphila JAB1、JAB1；Pseudomonas alcaliphila alcaligenes strain B2 16S、B2 16S；Pseudomonas pseudoalcaligenes strain C70c 16S、C70c 16S；Pseudomonas pseudoalcaligenes strain C70b 16S、C70b 16S；Pseudomonas pseudoalcaligenes strain C70a 16S，C70a 16S；Luteibactor rhizovicina isolate OUCZ70 16S、OUCZ70 16S；Luteibactor rhizovicinaDSM 16549、DSM 16549；Luteibactor rhizovicinastrain LJ96 16S、LJ96 16S；Bacillus cereus strain FSI-1 16S\u003c/em\u003e 、\u003cem\u003eFSI-1 16S\u003c/em\u003e ；\u003cem\u003eBacillus cereus strain GT36 16S、GT36 16S；Bacillus cereus strain MOB-5 16S、MOB-5 16S；Bacillus megaterium strain CS14 16S、CS14 16S；Bacillus megaterium strain CS4 16S、CS4 16S；Bacillus megaterium strain H2 16S、H2 16S\u003c/em\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/92b1ddd0b38eff84e2fdbb6b.png"},{"id":58260381,"identity":"400f5322-2868-4670-a525-b6107f3c7c7c","added_by":"auto","created_at":"2024-06-13 06:09:02","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":11033,"visible":true,"origin":"","legend":"\u003cp\u003eMicrobial count of ginseng rhizosphere soil under different forest types（ × 10\u003csup\u003e6 \u003c/sup\u003ecfu/g)\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/0eb65a28188c135ce6e781a6.png"},{"id":58260864,"identity":"1c1c37e2-3ca5-4e27-99cf-d4c5c05e29cb","added_by":"auto","created_at":"2024-06-13 06:17:02","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":16602,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Rhizosphere Soil Bacteria on Saponin Transformation in Panax ginseng under Different Forest Types (Unit: %)\u003c/p\u003e\n\u003cp\u003eFigure caption. Group I is the control group; Group II is the \u003cem\u003eBacillus pseudomycoides \u003c/em\u003egroup; Group III is the \u003cem\u003eBacillus subtilis\u003c/em\u003e group; Group IV is the \u003cem\u003ePseudomonas alcaliphila \u003c/em\u003egroup; Group V is a group of \u003cem\u003ePseudomonas pseudoalcaligenes \u003c/em\u003e; Group VI is the group of \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e; Group VII is the \u003cem\u003eBacillus cereus\u003c/em\u003e group; Group VIII is the \u003cem\u003eBacillus megaterium\u003c/em\u003e group; Group IX is the LF group (mixed of\u003cem\u003eBacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus megaterium\u003c/em\u003e); Group X is the CPF group (mixed with \u003cem\u003eBacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus cereus、Bacillus megaterium\u003c/em\u003e); Group XI is a BF group (mixed with \u003cem\u003eBacillus pseudomycoides、Bacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus megaterium\u003c/em\u003e); The XII group is the TH group (mixed with \u003cem\u003eBacillus pseudomycoides、Bacillus subtilis、Pseudomonas alcaliphila、Luteibactor rhizovicina、Bacillus megaterium\u003c/em\u003e).\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/f2ddc5c58087259edd60f190.png"},{"id":58260385,"identity":"335e96be-8f2d-461f-9938-7c655df93b3f","added_by":"auto","created_at":"2024-06-13 06:09:02","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":14260,"visible":true,"origin":"","legend":"\u003cp\u003eThe correlation coefficient between microorganisms and saponin content\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/0d02a11010c52716a414a0e6.png"},{"id":60113836,"identity":"0b81175c-cf28-42ba-8684-6cf42f87ac3e","added_by":"auto","created_at":"2024-07-12 02:49:37","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3915483,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/aab12510-5ca6-4e7b-afea-199d9e6ee102.pdf"},{"id":58260386,"identity":"4a8f9fd8-26d7-4d6f-8c5c-e98158288b4d","added_by":"auto","created_at":"2024-06-13 06:09:02","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":14345,"visible":true,"origin":"","legend":"","description":"","filename":"AppendixA.docx","url":"https://assets-eu.researchsquare.com/files/rs-4487770/v1/10fe6fd7fe92223ff184883d.docx"}],"financialInterests":"","formattedTitle":"Correlation analysis between ginsenoside content and rhizosphere soil microbial species in different forest types","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe rhizosphere microorganisms of plants are considered as the \"second genome of plants\"(Wang et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).The growth, development, yield, secondary metabolites, and disease defense of medicinal plants are crucial. The community structure and diversity of rhizosphere microorganisms are influenced by soil environment, which is related to factors such as forest type. Different forest types have different plant root exudates and accumulated humus, resulting in different soil environments(Man et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).Studying the rhizosphere microorganisms of medicinal plants under different forest types and their impact on the accumulation of effective components in medicinal plants is of great significance. \u003cem\u003ePanax ginseng\u003c/em\u003e C.A. Mey. is a perennial perennial herbaceous plant of the Panax genus in the Araliaceae family. It has a history of more than 5000 years of medicinal use in China and is commonly known as \"stick hammer\"(Potenza et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).It has traditional effects such as restoring meridians, strengthening the spleen and benefiting the lungs, and greatly nourishing vital energy(Jaiswal et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Plant chemistry studies have shown that the main active ingredient of ginseng is the triterpenoid compound ginsenoside(Kim et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Modern pharmacological studies have shown that ginsenosides have good anti-tumor, anti-inflammatory, antioxidant, and anti apoptotic effects(Gao et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).With the increasing awareness of health among people, the demand for ginseng is also gradually increasing. The cultivation methods of ginseng mainly include three types: farmland ginseng, garden ginseng, and mountainous forest cultivated ginseng(MFCG). MFCG, also known as cultivated mountain ginseng or seed sea, is divided into forest seed ginseng and forest transplanted ginseng. MFCG is a wild mountain ginseng that artificially grows ginseng seeds or seedlings in the mountains and forests. Compared with the other two cultivation methods of ginseng, the growth environment of ginseng under the forest is closer to that of wild mountain ginseng, and the quality of MFCG for more than ten years can be comparable to that of wild mountain ginseng. The content and type of ginsenosides are important indicators for evaluating ginseng. Although the yield and quality of ginsenosides have been widely studied, the yield of ginsenosides is still very low. Therefore, it is urgent to explore the cumulative impact mechanism of ginsenosides and improve the yield of ginsenosides.\u003c/p\u003e \u003cp\u003eThe cultivation history of ginseng is long, and China is the country with the largest ginseng planting area in the world(Potenza et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The Changbai Mountain area in the eastern part of Jilin Province is the main production area of ginseng in China. With the rapid development of the ginseng industry, the planting area of MFCG is also significantly increasing. Ecological factors such as forest type are the main influencing factors on the accumulation of ginsenosides in understory ginseng. Currently, research on the active ingredients of ginseng mainly focuses on pharmacological activity and quality control. However, in terms of microecology, there are currently few literature reports on the mechanism by which forest types affect the accumulation of ginsenosides. Therefore, studying the mechanism of different forest types affecting the accumulation of ginsenosides in MFCG is particularly important for improving the quality of ginseng.\u003c/p\u003e \u003cp\u003eThe ecosystems under different forest types are also different, and various ecological factors interact and interweave vertically and horizontally, forming a complex network. The ecological networks constructed by different tree species also have significant differences, such as the physical and chemical properties, microbial types and quantities, canopy closure, etc. of the soil under the forest(Bei et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), which leads to forest types becoming one of the important factors affecting the accumulation of ginsenosides. Studying the microbial effects of different forest types on the accumulation of ginsenosides is not only helpful for improving the quality of MFGG, but also crucial for agricultural development and ecological balance.\u003c/p\u003e \u003cp\u003eThe aim of this study is to reveal the specific differences in soil microbial activity and quantity among four forest types in the Changbai Mountain area: TH, LF, BF, and CPF. Dilution culture and top purification methods were used to isolate and identify soil microorganisms in the rhizosphere of ginseng under different forest types. At the same time, the content of 21 ginsenosides in ginseng was determined by high-performance liquid chromatography (HPLC). Through correlation analysis, the microbial accumulation mechanism of ginsenoside content influenced by forest type was revealed, thereby improving ginsenoside content. Provide theoretical basis for promoting the accumulation of effective ingredients in medicinal materials by rhizosphere microorganisms.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003cstrong\u003etudy site\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was conducted in the mountainous areas of Jingyu County, Jilin Province, including CPF, BF, LF, and TH(longitude 126 \u0026deg; 30 \u0026apos;-127 \u0026deg; 16\u0026apos;N, latitude 42 \u0026deg; 06\u0026apos; -42 \u0026deg; 48\u0026apos;E ，fig1.). Jingyu County is located in the hinterland of Changbai Mountain, with a forest coverage rate of up to 84%. It is the main production area of ginseng in Jilin Province and a representative area for studying forest ginseng. This mountainous area belongs to the Changbai Mountain Range. The mountainous area belongs to a continental monsoon climate in the cold temperate zone, with long and cold winters and short and warm and humid summers, with moderate sunlight. Altitude: 475 meters. Annual average temperature: 3.7 ℃, highest temperature is 37 \u0026deg;C, lowest temperature is -48 \u0026deg;C, annual average precipitation is 749.7 mm, and soil depth exceeds 25 centimeters. Table 1 summarizes the quality of the four forest types selected in this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1.\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eCharacteristics of four common planting forest types of understory ginseng in Jingyu County, Jilin Province.\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\"\u003e\u003c/p\u003e\n\u003cp\u003eNotes:TH，theropencedrymion ；LF，larch forest \u0026nbsp; ；BF，broad-leaved forest；CPF \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; camphor pine forest\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSample collection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNine 15-year-old disease-free MFCG from four different forest types were selected. The non-medicinal parts were removed, and the rhizomes were washed, dried, and ground into a fine powder. The ginsenoside content was then analyzed by crushing (0.425 sieve) the powder from each plant. Soils were collected separately from the four forest types mentioned above. Five sample points were selected in the same plot using the \u0026apos;S-type sampling method\u0026apos;. Soil humus was removed to a depth of 0-20 cm from the surface. The soil samples from the healthy ginseng rhizosphere under different forest types were collected uniformly, placed in sterile ziplock bags, and mixed to ensure representativeness. The samples were then quickly stored in an icebox and transported to the laboratory where they were immediately stored at -80◦C until microbial analysis was required.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetermination of ginsenoside content in\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eMFCG\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;samples\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe content of ginsenosides Rg1, Re, Rf, Rb1, Rg2, Rg3, Rg5, Rc, F1, Rb2, F2, Rh2, Rd, Rb3, compound K, 20 (R)-Rh1, Rk3, Rh4, original ginsenosides, and original ginsenosides under different forest types were determined by HPLC. Weigh 500 mg of ginseng powder for each of the four forest types. Place each sample in a 10 mL centrifuge tube and add 5.0 mL of chromatographic methanol solution. Seal the tubes, weigh them, and extract the samples using ultrasonication (40 Hz, 100 watts power, 45 min). The samples were left overnight, reweighed and sonicated for another 45 min, and the supernatant was transferred to a centrifuge tube with a sterile syringe and centrifuged at 5000 rpm for 15 min, and then filtered through a 0.22\u0026nbsp;\u0026mu;M\u0026nbsp;filter into a 2.0 mL sample vial for spare use. Weigh 5 mg each of ginsenoside standards Rg1, Re, Rg2, Rg3, Rg5, Rf, F1, F2, Rc, Rd, Rb1, Rb2, Rb3, Rh2, compound K, 20(R)-Rh1, Rk3, Rh4, protopanaxadiol and protopanaxatriol, and add methanol to formulate the mass concentrations of 1.04, 1.02, 0.98 , 1.00, 1.00, 1.02, 1.00, 1.00, 1.00, 1.02, 1.00, 1.00, 1.00, 1.04, 1.00, 1.00, 1.00, 0.98, 1.00, 1.00, 1.00, 1.02 mg/mL, and then filtrated through 0.45\u0026nbsp;\u0026mu;M\u0026nbsp;filtration membrane, and set aside.\u003c/p\u003e\n\u003cp\u003eThe chromatographic conditions refer to the method established in our laboratory for simultaneous determination of 20 types of ginsenosides.\u0026nbsp;The chromatographic column used in the study was BDS C\u003csub\u003e18\u003c/sub\u003e (250 mm \u0026times; 4.6 mm, 5 \u0026mu;m). The mobile phase was acetonitrile (A) and water (B) with the gradient elution as follows: 0-40 min, 18% A\u0026rarr; 21%; 40-42 min, 21% A\u0026rarr; 26%; 42-46 min, 26% A\u0026rarr; 32%; 46-66 min, 32% A\u0026rarr; 33.8%; 66-71 min, 33.8% A\u0026rarr; 38%; 71-77.7 min, 38% A\u0026rarr; 49.08%; 77.7-78 min, 49.08% A\u0026rarr; 49.1%; 78-82 min, 49.1% A; 82-83 min, 49.1% A\u0026rarr; 50.6%; 83-88 min, 50.6% A\u0026rarr; 59.6%; 88-89.8 min, 59.6% A\u0026rarr; 64.96%; 89.8-92 min, 64.96% A\u0026rarr; 65%; 92-97 min, 65% A; 97-102 min, 65% A\u0026rarr; 85%; 102-109 min, 85% A; 109-111 min, 85% A\u0026rarr; 18%. The velocity of the mobile phase was 1 ml/min. The detection wavelength was set as 203 nm while the temperature of the chromatographic column was 35 \u0026deg;C.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIsolation and purification of soil microorganisms in the rhizosphere of ginseng under different forest types\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe wet weight of ginseng inter-root soil samples under different forest types was accurately weighed 0.5 g, added with 5 mL of sterile water, and incubated at 28\u0026nbsp;℃\u0026nbsp;for 30 min with shaking, and then diluted according to the 10-fold dilution method, respectively, and 100\u0026nbsp;\u0026mu;L of bacterial suspension with a concentration of \u0026nbsp;1\u0026times;10\u003csup\u003e-7\u0026nbsp;\u003c/sup\u003ewas taken to coat the plate, and each concentration was repeated three times, and it was placed in the incubator at a constant temperature of 28\u0026nbsp;℃\u0026nbsp;for 1 d-3 d, and then it was reserved for use. The obtained colonies were isolated and purified according to different morphologies, and placed in an incubator at 28\u0026nbsp;℃\u0026nbsp;for inverted culture for 1-3d to observe and record the morphology of the colonies, and when the purification was complete, the colonies were transferred to the corresponding slant for preservation, and named according to their morphological characteristics.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIsolation and identification of soil microorganisms in the rhizosphere of ginseng under different forest types\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSoil bacteria and fungi DNA were extracted using Bacterial Genomic DNA Extraction Kit (Solarbio, Beijing China) and Fungi Genomic DNA Extraction Kit (Solarbio, Beijing China), respectively. The extracted DNA is purified and stored at -20℃ for future use. PCR amplification of bacterial 16s RNA was performed using bacterial universal primers 1492R and 27F. The primers were provided by Bao Bio-engineering Co., Ltd (Dalian, China) and the sequences were 5\u0026apos;-TACGGCTACCTTGTTACGACTT-3\u0026apos; and 5\u0026apos;-AGAGTTTGATCCTGGGCTCAG-3\u0026apos;. \u0026nbsp;The 20 \u0026mu;L PCR reaction system consists of 12.5 \u0026mu;L of 2\u0026times;Taq MasterMix for Page, 1 \u0026mu;L of upstream primer, 1 \u0026mu;L of downstream primer, 3 \u0026mu;L of DNA template, and 7.5 \u0026mu;L of deionized water. PCR reaction procedure was: pre denaturation at 95 ℃ for 5 min, denaturation at 95 ℃ for 30 s, then annealing at 57℃/54 ℃ for 30 s, and 30 cycles of extension at 72 ℃ for 90 s, and finally extension at 72 ℃ for 10 min. PCR products were detected by 1% agarose gel electrophoresis to observe whether there were specific target bands. The PCR products were then sequenced, and the sequencing service was provided by Sangon Biotech (Shanghai) Co., Ltd (Shanghai, China). The sequences were analyzed by Basic Local Alignment Search Tool (BLAST) in the National Center for Biotechnology Information (NCBI) database to identify the strains with the highest similarity to each other and to determine the biological classification status of each strain.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCorrelation analysis of ginseng inter-root soil microorganisms with ginsenosides under different forest types\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe number of each colony of soil under each forest type obtained above and the measured data of ginsenosides of ginseng under each forest type were entered into SPSS 22.0 (Chicago, USA) software for correlation analysis and Pearson Correlation values were obtained, respectively. GraphPad Prism 8.0.2 (San Diego, USA) was used to map the number of ginseng inter-root microorganisms and the content of active compounds under different forest types.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInfluence of ginseng inter-root microorganisms on the transformation of ginsenosides under different forest types\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe seven colonies after 24 h of streaking culture on solid potato dextrose agar (PDA) medium were transferred to conical flasks containing 100 mL of liquid PDA medium with a grafting ring for each single colony respectively, and were reciprocally cultured on a shaker at 28 ℃, 140 r/min for 24 h, and the cultures of \u003cem\u003eBacillus pseudomycoides,Bacillus subtilis\u003c/em\u003e,\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e,\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e, \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e, \u003cem\u003eBacillus cereus\u003c/em\u003e, and \u003cem\u003eBacillus megaterium\u0026nbsp;\u003c/em\u003ewere aseptically preserved and stored for spare parts.\u003c/p\u003e\n\u003cp\u003eWeigh 0.5 g of ginseng powder through 20 mesh Pharmacopoeia sieve in 36 test tubes, dry heat sterilize at 80 ℃ for 2 h, add 3 mL of sterile water to each test tube separately with pipette gun, and cool to room temperature. Under aseptic conditions, 2mL of each group of bacterial solution was separately pipetted into the above test tubes. The specific information of the groups and inoculated bacterial solution is shown in Table 2. The tubes were then placed in an incubator at 28 ℃ with shaking, and the changes in ginsenoside content were observed after 8 days of incubation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e Grouping and bacterial liquid information\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003eGroups\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003eBacterial solution\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003esterile water\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus pseudomycoides\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus pseudomycoides\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus subtilis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus subtilis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;Pseudomonas alcaliphila\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;Pseudomonas alcaliphila\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003e\u003cem\u003ePseudomonas pseudoalcaligene\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003ePseudomonas pseudoalcaligene\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003e\u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus cereus\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus cereus\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus megaterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus megaterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003eLarch forest\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus subtilis\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003ePseudomonas pseudoalcaligene\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eBacillus megaterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003eCamphor pine forest\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus subtilis\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003ePseudomonas pseudoalcaligene\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eBacillus cereus\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eBacillus megaterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003eBroad-leaved forest\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eBacillus subtilis\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003ePseudomonas pseudoalcaligene\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eBacillus megaterium\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"35.73943661971831%\"\u003e\n \u003cp\u003eTheropencedrymion\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"64.2605633802817%\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eBacillus subtilis\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eBacillus megaterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003cstrong\u003etatistic analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe obtained data will be statistically analyzed using SPSS 22.0 (Chicago, USA) software. One-way ANOVA was used to analyze the differences in ginsenoside content among different forest types. Mean values were analyzed using Tukey\u0026apos;s test. \u003cem\u003eP\u0026nbsp;\u003c/em\u003e\u0026lt; 0.05 was considered to indicate significant differences, and all data were expressed as mean \u0026plusmn; standard deviation.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eEffect of different forest types on ginsenoside content in ginseng\u003c/h2\u003e\n \u003cp\u003eThe ginseng under four forest types all contain 14 types of ginsenoside monomers, as shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e and Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The content of Rg1 and Rf, as well as the sum of 20 monomeric saponins, were significantly higher in the CPF group than in the other three forest types (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The content of Re, Rb2, and Rb3 in the CPF group was significantly higher than that in the LF group and the BF group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05), while the differences between the LH group and the other groups were not significant. The Rb1 content in the CPF group was significantly higher than that in the other three forest types (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05), while Rb1 content was higher (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in the TH and BF groups than in the LF group. The Rg2 content in the LF group was significantly higher than that in the other three forest types (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The Rd content in TH group and the CPF group was significantly higher than the other two groups (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The Rh4 content in the LF group was significantly higher than that in the TH group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The content of ginsenoside Rg5 in the CPF group and the BF group was significantly higher than the other two groups (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Under the CPF forest type, the saponin content in the ginseng under the forest is the highest.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eIsolation, purification, and morphological observation of soil microorganisms in the rhizosphere of ginseng under different forest types\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eAs shown in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e and Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, separate and purify soil microorganisms to obtain 7 bacterial strains.\u003c/p\u003e\n \u003cp\u003eJQ.GSRS-1: The colonies are white, rod-shaped, with round ends and arranged in a chain like manner.\u003c/p\u003e\n \u003cp\u003eJQ.GSRS-2: The colony is dirty white, elliptical to columnar in shape, with a rough and opaque surface that forms wrinkles.\u003c/p\u003e\n \u003cp\u003eJQ.GSRS-3: The colony is white, rod-shaped, and has flagella at the end, which can move.\u003c/p\u003e\n \u003cp\u003eJQ.GSRS-4: The colony is green with flagella, round in shape, with neat edges and a smooth surface.\u003c/p\u003e\n \u003cp\u003eJQ.GSRS-5: The colony is yellow in color, round in shape, raised, and sticky, making it easy to pick up.\u003c/p\u003e\n \u003cp\u003eJQ.GSRS-6: The colony is in the shape of ground glass, the spores are round, the surface is rough, flat, and irregular, forming a snowflake like shape.\u003c/p\u003e\n \u003cp\u003eJQ.GSRS-7: The colony is milky white, rod-shaped, and has a round end.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003eThe number of bacteria in the rhizosphere soil of ginseng under different forest types\u003c/h2\u003e\n \u003cp\u003eAs shown in Fig. 6 the statistical results of the number of microorganisms in the rhizosphere soil of different forest types of ginsengs showed that a total of 7 bacteria were isolated and identified in the rhizosphere soil of four different forest types of MFCG. Among them, \u003cem\u003eBacillus pseudomycoides\u003c/em\u003e and \u003cem\u003eBacillus cereus\u003c/em\u003e were not isolated and identified in the MFCG soil of the LF type;\u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e was not isolated and identified in the MFCG soil of CPF type; \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e and \u003cem\u003eBacillus cereus\u003c/em\u003e were not isolated and identified in the MFCG soil of the TH type. \u003cem\u003eBacillus subtilis\u003c/em\u003e,\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e, \u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e and \u003cem\u003eBacillus megaterium\u003c/em\u003e were detected in all four types of forest MFCG soils.\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e was isolated and identified in BF and TH, with a quantity of 0.33 in the BF(\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003eBacillus pseudomycoides\u003c/em\u003e in the TH group is 0.35༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), there was no significant difference in the number of \u003cem\u003eBacillus pseudomycoides\u003c/em\u003e between the two forest types.\u003c/p\u003e\n \u003cp\u003eThe number of \u003cem\u003eBacillus subtilis\u003c/em\u003e in BF is 5.01(\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003eBacillus subtilis\u003c/em\u003e in the LF is 4.67༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003eBacillus subtilis\u003c/em\u003e in the CPF is 5.59༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003eBacillus subtilis\u003c/em\u003e in TH is 6.34༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), there was no significant difference in the number of \u003cem\u003eBacillus subtilis\u003c/em\u003e among the four forest types.\u003c/p\u003e\n \u003cp\u003eThe number of \u003cem\u003ePseudomonas alcaligenes\u003c/em\u003e in BF is 3.57(\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003ePseudomonas alcaligenes\u003c/em\u003e in the LF is 0.69༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003ePseudomonas alcaligenes\u003c/em\u003e in the CPF is 2.35༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003ePseudomonas alcaligenes\u003c/em\u003e in the TH is 0.68༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the content of \u003cem\u003ePseudomonas alcaligenes\u003c/em\u003e is higher in BF than in the other three forest types. The number of \u003cem\u003ePseudomonas alcaligenes\u003c/em\u003e is similar and smaller in LF and TH than in CPF.\u003c/p\u003e\n \u003cp\u003e\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e was detected in three forest types: BF, LF, and CPF. The number of \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e in the LF group was 9.57(\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), higher than the BF group and the CPF group, with a content of 2.03༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g) \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e in the BF group, the content of \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e in the CPF group is 3.47༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g).\u003c/p\u003e\n \u003cp\u003eThe number of \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e in the CPF is 38.13(\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g, significantly higher than the other three forest types, with a population of 17.69 \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e in BF༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e in the LF is 12.03༈\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), the number of \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e in the TH is 17.35༈\u0026times; 10\u003csup\u003e6\u003c/sup\u003e cfu/g).\u003c/p\u003e\n \u003cp\u003eWax like \u003cem\u003eBacillus cereus\u003c/em\u003e was detected in the CPF group, with a quantity of 4.3333(\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), no \u003cem\u003eBacillus cereus\u003c/em\u003e was detected in the other three forest types.\u003c/p\u003e\n \u003cp\u003eThe number of \u003cem\u003eBacillus megaterium\u003c/em\u003e in the LF is 23.47(\u0026times;10\u003csup\u003e6\u003c/sup\u003e cfu/g), significantly higher than the other three forest type groups. The number of \u003cem\u003eBacillus megaterium\u003c/em\u003e in the BF is 18.05 (\u0026times; 10\u003csup\u003e6\u003c/sup\u003e cfu/g).The number of \u003cem\u003eBacillus megaterium\u003c/em\u003e in the CPF is 12.02 (\u0026times; 10\u003csup\u003e6\u003c/sup\u003e cfu/g), and in the TH, the number of \u003cem\u003eBacillus megaterium\u003c/em\u003e is 8.37 (\u0026times; 10\u003csup\u003e6\u003c/sup\u003e cfu/g).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003eThe effect of ginseng rhizosphere microorganisms on the transformation of ginsenosides under different forest types\u003c/h2\u003e\n \u003cp\u003eTo verify the effect of different fungal species in the rhizosphere of ginseng under different forest types on the content of ginsenosides, a control group was set up based on the 7 identified bacteria in the bacteria detected under different forest types, Set up control groups separately,\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e group;\u003cem\u003eBacillus subtilis\u003c/em\u003e group;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e group; \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e group; \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e group; \u003cem\u003eBacillus cereus\u003c/em\u003e group;\u003cem\u003eBacillus megaterium\u003c/em\u003e group.Set up LF groups according to different forest types(mixed of \u003cem\u003eBacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus megaterium\u003c/em\u003e),CPF group (mixed with \u003cem\u003eBacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus cereus、Bacillus megaterium\u003c/em\u003e);BF group (mixed with \u003cem\u003eBacillus pseudomycoides、Bacillus subtilis、Pseudomonas alcaliphila、Pseudomonas pseudoalcaligene、Luteibactor rhizovicina、Bacillus megaterium\u003c/em\u003e); TH group \u003cem\u003e(\u003c/em\u003emixed with \u003cem\u003eBacillus pseudomycoides、Bacillus subtilis、Pseudomonas alcaliphila、Luteibactor rhizovicina、Bacillus megaterium\u003c/em\u003e). The results of ginsenoside detection in each group are shown in Fig. \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e and the 7 isolated and identified bacteria have different promoting effects on the monomer saponins.\u003c/p\u003e\n \u003cp\u003eThe group of \u003cem\u003eBacillus pseudomycoides\u003c/em\u003e had a significant promoting effect on the monomer saponin Rc (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); The \u003cem\u003eBacillus subtilis\u003c/em\u003e group had a good promoting effect on the monomeric saponins Rg2, Rb2, and Rd (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); The promotion effect of the \u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e group on the monomer saponin Rb3, diol type saponin, and saponin addition value is more significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); The group of \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e showed a significant promoting effect on the monomer saponin Rh2 (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); The group of \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003ehad a significant promoting effect on the monomer saponins Rg2, Rb3, F2, Rh4, diol type saponins, triol type saponins, and saponin addition values (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); The promotion effect of \u003cem\u003eBacillus cereus\u003c/em\u003e group on monomeric saponins Rc and Rb2 was more significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); The \u003cem\u003eBacillus megaterium\u003c/em\u003e group also has a good promoting effect on monomeric saponins Rc and Rb2. The mixed microbial colonies in ginseng soil under LF and TH types have a good promoting effect on the monomeric saponins Rc, Rb3, and diol type saponins (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); The CPF group had a significant promoting effect on the monomer saponin Rh4 (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); The BF group has a significant promoting effect on the monomer saponin Rb3 and diol type saponins (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eThe correlation between the accumulation of ginsenosides in different forest types and the rhizosphere soil microorganisms of MFCG\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe correlation analysis results (fig.8) indicate that there is varying degrees of correlation between the content of different monomeric saponins and the number of microorganisms, with the highest positive correlation coefficient between monomeric saponin Rg1 and \u003cem\u003eBacillus subtilis\u003c/em\u003e (r=0.5987). The negative correlation coefficient between Rg1 and \u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e was the highest (r=-0.7603), and the negative correlation coefficients of other soil microorganisms were ranked as follows: \u003cem\u003eBacillus pseudomycoides strain\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus cereus\u0026nbsp;\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus megaterium\u003c/em\u003e\u0026gt;\u003cem\u003e\u0026nbsp;Pseudomonas pseudoalcaligenes.\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between monomer saponin Re and \u003cem\u003eBacillus subtilis\u003c/em\u003e is the highest (r=0.7820). The negative correlation coefficient between Re and \u003cem\u003ePseudomonas pseudoalcaligenes\u0026nbsp;\u003c/em\u003ewas the highest (r=-0.8717), and the negative correlation coefficients of other soil microorganisms were ranked as follows: \u003cem\u003eBacillus megaterium\u003c/em\u003e\u0026gt;\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e\u0026gt;\u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e\u0026gt;\u003cem\u003ePseudomonas alcaliphila.\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between the monomer saponin Rf and \u003cem\u003eBacillus subtilis\u003c/em\u003e was the highest (r=0.7820), and the negative correlation coefficient between Rf and \u003cem\u003eBacillus megaterium\u003c/em\u003e was the highest (r=-0.8530). The negative correlation coefficient of other soil microorganisms was ranked as follows: \u003cem\u003eBacillus pseudomycoides\u003c/em\u003e\u0026gt;\u0026nbsp;\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e\u0026gt;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus cereus.\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eThe single saponin Rb1 has the highest positive correlation coefficient with \u003cem\u003eBacillus subtilis\u003c/em\u003e (r=0.9510). The negative correlation coefficient between Rb1 and \u003cem\u003eBacillus megaterium\u003c/em\u003e was the highest (r=-0.9560), and the negative correlation coefficients of other soil microorganisms were ranked as follows: \u003cem\u003ePseudomonas pseudoalcaligenes\u0026nbsp;\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus cereus\u003c/em\u003e\u0026gt;\u003cem\u003ePseudomonas alcaligenes.\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eThe single saponin Rg2 has the highest positive correlation coefficient with \u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e(r=0.9736), while the other soil microorganisms have the following positive correlation coefficients: \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus subtilis\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus megaterium\u0026nbsp;\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus subtilis\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e. The negative correlation coefficient between Rg2 and \u003cem\u003ePseudomonas alcaliphila\u0026nbsp;\u003c/em\u003eis the highest (r=-0.5280).\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between the monomer saponin Rc and \u003cem\u003eBacillus megaterium\u0026nbsp;\u003c/em\u003ewas the highest (r=0.9630), and the positive correlation coefficient of other soil microorganisms was ranked as follows: \u003cem\u003eBacillus pseudomycoides\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus subtilis\u003c/em\u003e. The negative correlation coefficient between Rc and\u003cem\u003e\u0026nbsp;Luteibactor rhizovicina\u003c/em\u003e is the highest (r=-0.6602), and the negative correlation coefficient of other soil microorganisms is ranked as follows: \u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e\u0026gt;\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus megaterium\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between monomer saponin Rb2 and \u003cem\u003eBacillus subtilis\u003c/em\u003e is the highest (r=0.9450). The negative correlation coefficient between Rb2 and \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e was the highest (r=-0.7286), and the negative correlation coefficients of other soil microorganisms were ranked as follows: \u003cem\u003eBacillus megaterium\u003c/em\u003e\u0026gt;\u003cem\u003eLuteibactor rhizovicina\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e\u0026gt;\u003cem\u003ePseudomonas alcaliphila.\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between monomeric saponin Rb3 and \u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e was the highest (r=0.9710), and the positive correlation coefficient of other soil microorganisms was ranked as follows: \u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus subtilis\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus cereus\u003c/em\u003e. The negative correlation coefficient between Rb3 and \u003cem\u003ePseudomonas pseudoalcaligenes\u0026nbsp;\u003c/em\u003ewas the highest (r=-0.7374), and the negative correlation coefficient of other soil microorganisms was ranked as follows: \u003cem\u003eBacillus megaterium\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between the monomer saponin Rd and \u003cem\u003eBacillus cereus\u0026nbsp;\u003c/em\u003ewas the highest (r=0.5722), and the positive correlation coefficient of other soil microorganisms was ranked as follows: \u003cem\u003eBacillus subtilis\u003c/em\u003e\u0026gt;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e\u0026gt;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e. Rd has the highest negative correlation coefficient with \u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e（r=-0.6132）,and the negative correlation coefficient of other soil microorganisms was ranked as follows: \u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e \u0026gt;\u003cem\u003eBacillus megaterium.\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between Rh4 and\u003cem\u003e\u0026nbsp;Luteibacter rhizovicinus\u003c/em\u003e was the largest (r=0.9590), and the positive correlation coefficients of other soil microorganisms were in the order of \u0026nbsp;\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e \u0026gt;\u003cem\u003e\u0026nbsp;Bacillus megaterium\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e. Rh4 had the largest negative correlation coefficient with\u0026nbsp;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e (r=-0.7875), and the other soil microorganisms were in the order of \u0026nbsp;\u003cem\u003eBacillus subtilis\u0026nbsp;\u003c/em\u003e\u0026gt;\u0026nbsp;\u003cem\u003eBacillus cereus\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between Compound K and\u0026nbsp;\u003cem\u003ePseudomonas alcaliphila\u0026nbsp;\u003c/em\u003ewas the largest (r=0.7413), and the positive correlation coefficients of other soil microorganisms were in the order of\u0026nbsp;\u003cem\u003eBacillus megaterium\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e. Compound K had the largest negative correlation coefficient with\u0026nbsp;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e (r=-0.7863), and the other soil microorganisms were in the order of\u0026nbsp;\u003cem\u003e\u0026nbsp;Bacillus subtilis\u0026nbsp;\u003c/em\u003e\u0026gt;\u0026nbsp;\u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus cereus\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between Rg5 and\u0026nbsp;\u003cem\u003eLuteibacter rhizovicinus\u0026nbsp;\u003c/em\u003ewas the highest (r=0.7699). The negative correlation coefficient between Rg5 and\u0026nbsp;\u003cem\u003eBacillus pseudomycoides\u0026nbsp;\u003c/em\u003ewas the largest (r=-0.9730), and the negative correlation coefficients of other soil microorganisms were in the order of\u0026nbsp;\u003cem\u003eBacillus megaterium\u0026nbsp;\u003c/em\u003e\u0026gt;\u0026nbsp;\u003cem\u003eBacillus cereus\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003ePseudomonas pseudoalcaligenes\u0026nbsp;\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between Rh2 and\u0026nbsp;\u003cem\u003ePseudomonas pseudoalcaligenes \u0026nbsp;\u003c/em\u003ewas the largest (r=0.8818), and the positive correlation coefficients of other soil microorganisms were in the order of\u0026nbsp;\u003cem\u003eBacillus megaterium\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e. Rh2 had the largest negative correlation coefficient with\u0026nbsp;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e (r=-0.7088), and the other soil microorganisms were in the order of \u0026nbsp;\u003cem\u003eBacillus subtilis\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus cereus\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between propanaxanediol and\u0026nbsp;\u003cem\u003eBacillus subtilis\u0026nbsp;\u003c/em\u003ewas the largest (r=0.8496). The negative correlation coefficient between propanaxanediol and\u0026nbsp;\u003cem\u003eBacillus megaterium\u003c/em\u003e was the largest (r=-0.8784), and the negative correlation coefficients of other soil microorganisms were in the order of\u0026nbsp;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003ePseudomonas alcaliphila\u0026nbsp;\u003c/em\u003e\u0026gt;\u0026nbsp;\u003cem\u003eBacillus cereus\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between diol type and\u0026nbsp;\u003cem\u003ePseudomonas alcaliphila\u0026nbsp;\u003c/em\u003ewas the largest (r=0.9620). The negative correlation coefficient between diol type and\u0026nbsp;\u003cem\u003eBacillus megaterium\u003c/em\u003e was the largest (r=-0.8304), and the negative correlation coefficients of other soil microorganisms were in the order of\u0026nbsp;\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e\u0026gt;\u0026nbsp;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus cereus\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between triol type and\u0026nbsp;\u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e was the largest (r=0.9339), and the positive correlation coefficient of other soil microorganisms was in the order of\u0026nbsp;\u003cem\u003ePseudomonas pseudoalcaligenes\u0026nbsp;\u003c/em\u003e\u0026gt;\u0026nbsp;\u003cem\u003eBacillus subtilis\u003c/em\u003e. The negative correlation coefficient between triol type and\u0026nbsp;\u003cem\u003eBacillus cereus\u0026nbsp;\u003c/em\u003ewas the largest (r=-0.6669), and the negative correlation coefficient of other soil microorganisms was in the order of\u0026nbsp;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus megaterium\u003c/em\u003e.\u003c/p\u003e\n \u003cp\u003eThe positive correlation coefficient between the addition value of monomonins and\u0026nbsp;\u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e was the largest (r=0.9633). The negative correlation coefficient between the sum of monomonins and\u0026nbsp;\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e was the largest (r=-0.6015), and the other negative correlation coefficients of soil microorganisms were in the order of\u003cem\u003e\u0026nbsp;Bacillus pseudomycoides\u003c/em\u003e \u0026gt;\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e \u0026gt;\u0026nbsp;\u003cem\u003eBacillus cereus\u003c/em\u003e.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eGinseng (\u003cem\u003ePanax ginseng\u0026nbsp;\u003c/em\u003eC.A. Meyer), as a representative rare traditional medicinal plant, has been eaten in China for more than 5000 years(Potenza et al. 2023). At present, studies have proved that ginseng has a good therapeutic effect on fatigue, anorexia, shortness of breath, palpitations, insomnia and diabetes(Gao et al. 2018). Due to its remarkable pharmacological properties, ginseng has been widely recognized in Asia and attracted a large number of consumer groups, but excessive mining has also led to the rapid extinction of wild ginseng resources in just a few decades(L 2015). In order to meet the increasing market demand, people began to widely plant ginseng under the forest to replace wild ginseng.\u003c/p\u003e\n\u003cp\u003eMFCG is a semi wild ginseng cultivated by artificially sowing ginseng seedlings or seeds under natural or artificial forests, simulating the growth process of wild ginseng. Its appearance and composition are similar to some characteristics of wild ginseng. China has been trying to cultivate ginseng under the forest in the Northeast region since the 1980s(F 2019). In recent years, in accordance with the requirements of China's ecological civilization construction policy regarding the development of understory economy, promoting understory ginseng cultivation has become the main trend in the development of ginseng cultivation in Northeast China. By relying on a series of industrial support policies from the government for the cultivation of ginseng under forests, the scale of ginseng cultivation in China has been further expanded(Liu et al. 2016).\u003c/p\u003e\n\u003cp\u003eThe pharmacological effects of ginseng are closely related to its multiple active ingredients in the body, including ginsenosides, polysaccharides, peptides, plant sterols, etc. Among them, ginsenosides are the main pharmacological active ingredients of ginseng. Ginsenoside belongs to the triterpenoid class of compounds, which are a type of terpenoid compounds composed of 30 carbon atoms in the basic nucleus, and are polymerized from 6 isoprene units. There are currently over 100 known types of ginsenosides, which can be divided into tetracyclic triterpenes and pentacyclic triterpenes based on their different mother ring structures. Ginsenoside has high medicinal activity and its pharmacological effects have been widely studied in recent years. The research results indicate that it has the effects of enhancing physical strength, improving immunity, and prolonging life. At the same time, it can also resist a variety of adverse reactions, including depression, diabetes, fatigue, aging, inflammation, tumors, lung problems, indigestion, vomiting, tension, stress, ulcers and other adverse reactions(Wang et al. 2009).\u003c/p\u003e\n\u003cp\u003eBased on the rich pharmacological activity of ginsenosides, how to improve the content of ginsenosides in ginseng medicinal materials has always been a hot research topic for scholars. However, due to the complex transformation and accumulation mechanism of ginsenosides in ginseng, there is still no good solution to this problem. The main factors affecting the accumulation of ginsenosides in MFCG are ecological factors such as forest type(Wang et al. 2021). However, the specific ways in which different forest types regulate the content of ginsenosides in MFCG have not been thoroughly studied. In recent years, an increasing number of studies have shown that soil rhizosphere microorganisms have a significant impact on the increase of effective ingredient content in medicinal plants(Wang et al. 2021). However, there has been no in-depth research on whether the rhizosphere soil microorganisms of Panax ginseng in different forest types are involved in the transformation and accumulation of ginsenosides in Panax ginseng. In this study, rhizosphere soil and MFCG samples were collected from four different forest types (CPF, BF, LF, and TH) in the mountainous area of Jingyu County, Jilin Province, China. Subsequently, the content of 20 ginsenosides in the MFCG samples under the forest was analyzed using high-performance liquid chromatography, and the microorganisms in the ginseng rhizosphere soil were isolated, purified, and identified. Based on the experimental results, we conducted a series of correlation analyses, including: correlation analysis between forest type and rhizosphere soil microorganisms, correlation analysis between rhizosphere soil microorganisms and ginsenoside transformation in ginseng under the forest, correlation analysis between rhizosphere soil microorganisms and ginsenoside accumulation in ginseng under the forest, etc.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffects of different forest types on MFCG quality\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp;Therefore, we selected several representative forest types planted under the forest type of understory ginseng as the research object, and determined the content of 20 important ginseng monomer saponins (Rg1, Re, Rf, Rb1, Rg2, Rc, Rh1, Rb2, Rb3, F1, Rd, Rk3, F2, Rh4, Rg3, There were 14 ginsenosides (Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, Rd, Rh4, Compound K, Rg5, Rh2, Progindadiol) in ginseng under the four forest types, but there were some differences in the content of monomeric saponins and the addition value of 14 saponins. Among them, the sum value of 14 monomeric saponins in Pinus sylvestris var. mongolica was the highest, and the quality was the best, so we believe that CPF was the most suitable for planting ginseng. The sum value of the 14 monomeric saponins of ginseng under the BF was the lowest.However, there was no change in the monomeric saponin species，This may be related to G × E (genotype × environment) interactions.For example, the activity of dogwood, basil, ashwagandha and notoginseng(Liu et al. 2018; Xu et al. 2023; Kumar et al. 2023).In addition, widely distributed species in order to adapt to different environments lead to the creation of stable genetic variation, resulting in multiple genotypes of the same species. It is possible that the composition and chemical content of these genotypes will be different, for example, trans-anethole, the main ingredient in fennel, is higher in both genotypes and there is no difference between the other genotypes(Yaldiz and Camlica 2019).We hypothesized that environmental variability may be one of the factors contributing to the significant differences in the content of ginsenosides, the main active ingredient of ginseng. This is also confirmed by other people's reports(Zhu et al. 2022).Indicates that differences in environmental conditions, as well as soil properties, may lead to substantial differences in the quality of medicinal plant material.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffects of different forest types on soil microorganisms in the rhizosphere of ginseng\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMicroorganisms in the soil play the role of decomposers in the forest ecosystem, and participate in the process of information transmission and energy flow in the forest(Li et al. 2021; Kunito et al. 2012).A total of 7 bacteria (\u003cem\u003eBacillus pseudomycoides\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003e\u0026nbsp;Pseudospora subtilis\u003c/em\u003e、\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003ePseudomonas pseudoalcaligenes\u0026nbsp;\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eLuteibacter rhizovicinus\u0026nbsp;\u003c/em\u003e\u003cem\u003e、\u003c/em\u003e\u003cem\u003eBacillus cereus\u003c/em\u003e and\u003cem\u003e\u0026nbsp;Bacillus megaterium\u003c/em\u003e) were isolated and identified in the rhizosphere soil of ginseng in four different forest types, among which the number of microorganisms was the highest in the BF soil, the highest number of \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003ein the pine of CPF, and there was no significant difference in the number of colonies in the LF and the TH. The results showed that there was a strong correlation between forest type and soil microorganisms, which was consistent with the results of previous studies(Liu et al. 2023).Soil microorganisms are closely related to the synthesis of ginsenosides, and Wang et al. studied that Aspergillus niger fungi can promote the production of ginsenosides(Wang et al. 2019).There is also Penicillium YJM-2013 that promotes the accumulation of ginsenosides by enhancing the production of signaling molecules, activating the expression of transcription factors and functional genes(Wang et al. 2020).Four bacteria (\u003cem\u003ePseudospora subtilis\u003c/em\u003e,\u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e, \u003cem\u003eLuteibacter rhizovicinus Bacillus megamegasporus\u003c/em\u003e) were detected in four different forest types.\u003cem\u003eBacillus cereus\u0026nbsp;\u003c/em\u003ewas detected only in the rhizosphere soil of ginseng under CPF.We found that therefore, we believe that forest type has a greater impact on the species and number of soil bacteria in the rhizosphere of ginseng. Therefore, we believe that the number and species of rhizosphere microorganisms in the soil of ginseng under the forest type were affected by the forest type, and \u003cem\u003eBacillus cereus\u003c/em\u003e promoted the accumulation of ginsenosides.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffects of rhizosphere soil microorganisms on medicinal plants\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCorrelation coefficient between ginsenoside accumulation and soil microorganisms in the rhizosphere of ginseng according to different forest types (fig. 8),It was found that the content of ginsenosides was more closely related to the types and quantities of soil bacteria, indicating that they played an important role in the formation of ginseng quality. Soil rooting bacteria affect the colonization of plant roots by growth-promoting bacteria and stimulate plant growth and development. Recent studies have proved that rhizosphere bacteria and secondary metabolites not only affect plant growth and resistance to stress stress, but also affect the absorption of nutrients and the accumulation of active ingredients in medicinal plants(Yang et al. 2009; Dhungana et al. 2023; Shang et al. 2023).For example, preliminary experiments in our laboratory have confirmed that \u0026nbsp;\u003cem\u003eBacillus cereus\u0026nbsp;\u003c/em\u003ehave a disease prevention and growth promotion effect on ginseng(Y 2023).Relevant representative studies have shown that moss and \u003cem\u003eBacillus subtilis\u003c/em\u003e can significantly increase the content and yield of artemisinin(Awasthi et al. 2011).Trichosa can increase the production of ginsenosides through biological induction(Xu et al. 2021).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; Therefore, we speculated that ginseng under different forest types may selectively recruit specific microorganisms to metabolize certain substances, thereby affecting ginsenoside content.Our experimental results showed that seven different bacteria were isolated from the four forest types.The enrichment of seven bacteria, including \u003cem\u003eBacillus pseudomycoides, Bacillus subtilis,Pseudomonas alcaliphila,Pseudomonas pseudoalcaligenes, Luteibacter rhizovicinus Bacillus cereus, and Bacillus megaterium,\u0026nbsp;\u003c/em\u003emay promote the biosynthesis and accumulation of ginsenosides, the triterpenoid glycoside compounds of ginseng.The mechanism of the effect of different forest types on microbial-mediated ginsenoside accumulation was clarified.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contributions \u0026nbsp;\u003c/strong\u003eConceptualization;Yugang Gao. Funding acquisition;Yugang Gao. Investigation; Fengyu Pang, Xiaojia Ruan, Qun Liu. Methodology;Yugang Gao, Yan Zhao, Fengyu Pang, Qun Liu. Supervision; Yugang Gao,Yan Zhao. Visualization; Fengyu Pang. Roles/Writing - original draft; Fengyu Pang, Xiaojia Ruan.writing - review \u0026amp; editing; Fengyu Pang, Xiaojia Ruan, Yan Zhao, Yugang Gao,Qun Liu.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding \u0026nbsp;\u003c/strong\u003eWe thank funding supports for current research from National Key Research and Development Programme (Grant No. 2022YFF1300503) and Science and Technology Development Program of Jilin Province (Grant No. 20220401110YY).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability \u0026nbsp;\u003c/strong\u003eData will be made available on request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e The authors declare no conflict of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAwasthi A, Bharti N, Nair P, Singh R, Shukla AK, Gupta MM, Darokar MP, Kalra A (2011) Synergistic effect of Glomus mosseae and nitrogen fixing Bacillus subtilis strain Daz26 on artemisinin content in Artemisia annua L. Applied Soil Ecology 49:125-130. doi:https://doi.org/10.1016/j.apsoil.2011.06.005\u003c/li\u003e\n\u003cli\u003eBei Q, Yang T, Ren C, Guan E, Dai Y, Shu D, He W, Tian H, Wei G (2023) Soil pH determines arsenic-related functional gene and bacterial diversity in natural forests on the Taibai Mountain. Environmental Research 220:115181. doi:https://doi.org/10.1016/j.envres.2022.115181\u003c/li\u003e\n\u003cli\u003eDhungana I, Kantar MB, Nguyen NH (2023) Root exudate composition from different plant species influences the growth of rhizosphere bacteria. Rhizosphere 25:100645. doi:https://doi.org/10.1016/j.rhisph.2022.100645\u003c/li\u003e\n\u003cli\u003eF H (2019) Transeriptional analysis of terpenoids metabolism in developing Panax ginseng root Beijing Forestry University\u003c/li\u003e\n\u003cli\u003eGao Y, Liu J, Ji Q, Zhao Y, Zang P, He Z, Zhu H, Zhang L (2018) Anti-tumor activity and related mechanism study of Bacillus Polymyxa transformed Panax ginseng C. A. Mey. Process Biochemistry 72:198-208. doi:https://doi.org/10.1016/j.procbio.2018.06.013\u003c/li\u003e\n\u003cli\u003eJaiswal Y, Liang Z, Zhao Z (2016) Botanical drugs in Ayurveda and Traditional Chinese Medicine. Journal of Ethnopharmacology 194:245-259. doi:https://doi.org/10.1016/j.jep.2016.06.052\u003c/li\u003e\n\u003cli\u003eKim JH, Yi Y-S, Kim M-Y, Cho JY (2017) Role of ginsenosides, the main active components of Panax ginseng, in inflammatory responses and diseases. Journal of Ginseng Research 41 (4):435-443. doi:https://doi.org/10.1016/j.jgr.2016.08.004\u003c/li\u003e\n\u003cli\u003eKumar A, Venugopal S, Jnanesha AC, Lal RK (2023) Agricultural-based challenges, genetic enhancement, and obstacles to an industrially important medicinal plant, ashwagandha (Withania somnifera (L.) Dunal): A review. Ecological Genetics and Genomics 28:100183. doi:https://doi.org/10.1016/j.egg.2023.100183\u003c/li\u003e\n\u003cli\u003eKunito T, Tobitani T, Moro H, Toda H (2012) Phosphorus limitation in microorganisms leads to high phosphomonoesterase activity in acid forest soils. Pedobiologia 55 (5):263-270. doi:https://doi.org/10.1016/j.pedobi.2012.05.002\u003c/li\u003e\n\u003cli\u003eL X (2015) Research on Prediction and Evaluation of Light Environment of Cultivated Ginseng under Forest. Jilin University\u003c/li\u003e\n\u003cli\u003eLi J, Zhu T, Singh BK, Pendall E, Li B, Fang C, Nie M (2021) Key microorganisms mediate soil carbon-climate feedbacks in forest ecosystems. Science Bulletin 66 (19):2036-2044. doi:https://doi.org/10.1016/j.scib.2021.03.008\u003c/li\u003e\n\u003cli\u003eLiu C, Zheng C, Wang L, Zhang J, Wang Q, Shao S, Qin H, Xu Q, Liang C, Chen J (2023) Moso bamboo invasion changes the assembly process and interactive relationship of soil microbial communities in a subtropical broadleaf forest. Forest Ecology and Management 536:120901. doi:https://doi.org/10.1016/j.foreco.2023.120901\u003c/li\u003e\n\u003cli\u003eLiu W-w, Liu M-c, Li W-h, Zeng F-s, Qu Y (2016) Influence of ginseng cultivation under larch plantations on plant diversity and soil properties in Liaoning Province, Northeast China. Journal of Mountain Science 13 (9):1598-1608. doi:10.1007/s11629-015-3753-x\u003c/li\u003e\n\u003cli\u003eLiu Y-R, Delgado-Baquerizo M, Wang J-T, Hu H-W, Yang Z, He J-Z (2018) New insights into the role of microbial community composition in driving soil respiration rates. Soil Biology and Biochemistry 118:35-41. doi:https://doi.org/10.1016/j.soilbio.2017.12.003\u003c/li\u003e\n\u003cli\u003eMan H, Dong X, Li M, Zheng Z, Wang C, Zang S (2023) Spatial distribution and influencing factors of humus layer thickness of forest land in permafrost region of Northeast China. CATENA 224:106979. doi:https://doi.org/10.1016/j.catena.2023.106979\u003c/li\u003e\n\u003cli\u003ePotenza MA, Montagnani M, Santacroce L, Charitos IA, Bottalico L (2023) Ancient herbal therapy: A brief history of Panax ginseng. Journal of Ginseng Research 47 (3):359-365. doi:https://doi.org/10.1016/j.jgr.2022.03.004\u003c/li\u003e\n\u003cli\u003eShang X-c, Zhang M, Zhang Y, Hou X, Yang L (2023) Waste seaweed compost and rhizosphere bacteria Pseudomonas koreensis promote tomato seedlings growth by benefiting properties, enzyme activities and rhizosphere bacterial community in coastal saline soil of Yellow River Delta, China. Waste Management 172:33-42. doi:https://doi.org/10.1016/j.wasman.2023.09.003\u003c/li\u003e\n\u003cli\u003eWang B, Chen C, Xiao Y-M, Chen K-Y, Wang J, Zhao S, Liu N, Li J-N, Zhou G-Y (2024) Trophic relationships between protists and bacteria and fungi drive the biogeography of rhizosphere soil microbial community and impact plant physiological and ecological functions. Microbiological Research 280:127603. doi:https://doi.org/10.1016/j.micres.2024.127603\u003c/li\u003e\n\u003cli\u003eWang CQ, Yi LW, Zhao L, Zhou YZ, Guo F, Huo YS, Zhao DQ, Xu F, Wang X, Cai SQ (2021) 177 Saponins, Including 11 New Compounds in Wild Ginseng Tentatively Identified via HPLC-IT-TOF-MS(n), and Differences among Wild Ginseng, Ginseng under Forest, and Cultivated Ginseng. Molecules 26 (11). doi:10.3390/molecules26113371\u003c/li\u003e\n\u003cli\u003eWang S-h, Liang W-x, Lu J, Yao L, Wang J, Gao W-y (2020) Penicillium sp. YJM-2013 induces ginsenosides biosynthesis in Panax ginseng adventitious roots by inducing plant resistance responses. Chinese Herbal Medicines 12 (3):257-264. doi:https://doi.org/10.1016/j.chmed.2020.02.003\u003c/li\u003e\n\u003cli\u003eWang S, Wang J, Liang W, Yao L, Gao W (2019) Promotion of ginsenosides production in a co-cultivation system of Panax ginseng adventitious roots and immobilized Aspergillus niger. Industrial Crops and Products 140:111564. doi:https://doi.org/10.1016/j.indcrop.2019.111564\u003c/li\u003e\n\u003cli\u003eWang Y-Z, Chen J, Chu S-F, Wang Y-S, Wang X-Y, Chen N-H, Zhang J-T (2009) Improvement of Memory in Mice and Increase of Hippocampal Excitability in Rats by Ginsenoside Rg1\u0026rsquo;s Metabolites Ginsenoside Rh1 and Protopanaxatriol. Journal of Pharmacological Sciences 109 (4):504-510. doi:https://doi.org/10.1254/jphs.08060FP\u003c/li\u003e\n\u003cli\u003eXu X-d, Liang W-x, Yao L, Paek K-Y, Wang J, Gao W-y (2021) Production of ginsenoside by Chaetomium sp. and its effect on enhancing the contents of ginsenosides in Panax ginseng adventitious roots. Biochemical Engineering Journal 174:108100. doi:https://doi.org/10.1016/j.bej.2021.108100\u003c/li\u003e\n\u003cli\u003eXu Y, Zhu M, Feng Y, Xu H (2023) Panax notoginseng-microbiota interactions: From plant cultivation to medicinal application. Phytomedicine 119:154978. doi:https://doi.org/10.1016/j.phymed.2023.154978\u003c/li\u003e\n\u003cli\u003eY T (2023) The effect of symbiotic bacteria and trace elements on the degradation of pesticide residues by Bacillus polymyxoides. Jilin Agricultural University\u003c/li\u003e\n\u003cli\u003eYaldiz G, Camlica M (2019) Variation in the fruit phytochemical and mineral composition, and phenolic content and antioxidant activity of the fruit extracts of different fennel (Foeniculum vulgare L.) genotypes. Industrial Crops and Products 142:111852. doi:https://doi.org/10.1016/j.indcrop.2019.111852\u003c/li\u003e\n\u003cli\u003eYang J, Kloepper JW, Ryu C-M (2009) Rhizosphere bacteria help plants tolerate abiotic stress. Trends in Plant Science 14 (1):1-4. doi:https://doi.org/10.1016/j.tplants.2008.10.004\u003c/li\u003e\n\u003cli\u003eZhu L, Xu L, Huang Y, Xie C, Dou D, Xu J (2022) Correlations between ecological factors and the chemical compositions of mountainous forest cultivated ginseng. Journal of Food Composition and Analysis 114:104867. doi:https://doi.org/10.1016/j.jfca.2022.104867\u003c/li\u003e\n\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":"Forest types, Ginseng, Saponin accumulation, Soil microorganism","lastPublishedDoi":"10.21203/rs.3.rs-4487770/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4487770/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eInter-root microorganisms are major factors influencing the formation of high-quality herbs and promoting the accumulation of secondary metabolites, and this relationship has been well characterised in many supra-herbal medicinal herbs, but there is limited information on whether ginseng saponin accumulation is mediated by microorganisms in different forest types.Inter-root soil samples of ginseng and ginseng samples were collected from four different forest types (Theropencedrymion, Larch forest, Broad-leaved forest and Camphor pine forest) in the mountainous areas of Jingyu County, Jilin Province, China. The content of ginsenosides in the collected ginseng samples was determined by high performance liquid chromatography (HPLC). The results showed that the content of ginsenosides in Camphor pine forest was significantly higher than that in the other three forest types.The microorganisms in the soil samples were isolated and purified, and subsequently sequenced and analyzed by high-throughput sequencing methods, and a total of seven bacterial species were isolated and identified in the inter-root soil of ginseng from four different forest types. In broad-leaved forests (BF) and larch forests (LF), \u003cem\u003eBacillus megaterium\u003c/em\u003e is the most abundant microorganism. In the camphor pine forests (CPF) and theropencedrymion (TH), \u003cem\u003eLuteibactor rhizovicina\u003c/em\u003eis the largest proportion of microorganisms.\u003cstrong\u003e \u003c/strong\u003eRelevant analysis shows that several identified strains from the four forest types, including \u003cem\u003eBacillus pseudomycoides\u003c/em\u003e, \u003cem\u003eBacillus subtilis\u003c/em\u003e, \u003cem\u003ePseudomonas alcaliphila\u003c/em\u003e, \u003cem\u003eLuteibacter rhizovicinus\u003c/em\u003e and \u003cem\u003ePseudomonas alcaliphila\u003c/em\u003ecan promote the biosynthesis and accumulation of monomeric saponins Rc, Rb1, Rb2, Rb3, Rg2, Rb3, and Rh4. Our research findings emphasize the crucial role of different forest stand types in soil microbial community structure, and explore the accumulation mechanism of ginsenosides from a microbial perspective. In summary, this study provides more theoretical basis for the relationship between different forest types and the bioactive components of medicinal plants.\u003c/p\u003e","manuscriptTitle":"Correlation analysis between ginsenoside content and rhizosphere soil microbial species in different forest types","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-13 06:08:57","doi":"10.21203/rs.3.rs-4487770/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":"40be70f9-94c7-4fa3-bb0a-6ccf92dab0ea","owner":[],"postedDate":"June 13th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-07-12T02:41:29+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-13 06:08:57","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4487770","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4487770","identity":"rs-4487770","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}