Pseudostellaria Heterophylla Research Articles

Pseudostellaria heterophylla cultivar mixtures driven changes in rhizosphere metabolites to suppress soil-borne Fusarium disease

Crop diversification contributes to a decrease in soil-borne crop diseases, as well as an increase in agricultural productivity. However, few studies have investigated the changes in the composition of the rhizosphere microbial communities and rhizosphere metabolites, as well as their suppressive effect on soil-borne diseases under different crop cultivar mixture regimes. We carried out a series of experiments to assess changes in the rhizosphere microbial community and metabolites profile under different Pseudostellaria heterophylla cultivar mixture cultivation in consecutive monoculture fields by employing amplicon metagenomics (16S rRNA, ITS, and 18S rRNA) and non-targeted metabolomics. The impact of key metabolites on pathogenic Fusarium oxysporum, crop growth, and soil microorganisms was assessed under controlled conditions. Our study indicated that the cultivar mixtures improved the P. heterophylla performance, increased the fresh root biomass by 81.9–115.4 % and the heterophyllin B content by 35 % compared to the consecutive monoculture, respectively. Cultivar mixtures increased the abundance of beneficial bacteria (Lactobacillus, Pseudomonas, Nitrosospira) and consumer protists, and decreased the abundance of pathogenic fungal genera (Fusarium, Alternaria, Curvularia, Stemphylium, Gibberella). The qPCR results indicated that the cultivar mixtures significantly decreased the abundance of pathogenic F. oxysporum by 64.0–84.3 % compared to the consecutive monoculture treatment. Non-targeted metabolomics analysis showed that the cultivar mixtures significantly altered the soil metabolite profiles, and increased the contents of d-galactose, galactinol, d-sorbitol, glycerol, melibiose, D-fructose and D-tagatose. Subsequently, the key upregulated metabolites (glycerol, d-fructose, gluconic acid, quinic acid, and l-valine), identified through the random forest analysis, significantly inhibited the growth of F. oxysporum. The crucial metabolites in the presence of a pathogen (F. oxysporum) and single metabolite treatment significantly increased the biomass, SOD and CAT activity and decreased the POD and MAD activity of P. heterophylla compared to FOP (F. oxysporum treatment). Furthermore, the crucial metabolites under pathogen treatment significantly lowered the abundance of total fungi and F. oxysporum and increased the abundance of Pseudomonas spp. compared to FOP. Therefore, our study was able to emphasize the efficacy of using cultivar mixtures to combat soil-borne Fusarium disease through the modulation of rhizosphere metabolites.

Correlation analysis between Radix Pseudostelariae quality and rhizosphere soil factors of Pseudostellaria heterophylla in a cultivation region, China.

Pseudostellaria heterophylla, known for its significant bioactive ingredients, offers potential health benefits. Amounts of bioactive compounds of the tuberous root of cultivated Pseudostellaria heterophylla are sensitive to environmental conditions. We selected 22 sampling sites in Guizhou Province, China, a primary Pseudostellaria heterophylla planting area. We analyzed polysaccharides, water-soluble extractives, total ash and inorganic elements (Fe, Mn, Zn, Mg and Ca) in Radix Pseudostellariae, and pH, organic carbon (OC), available nitrogen (AN), available phosphorus (AP), available potassium (AK) and inorganic elements in the soil. Our study revealed a substantial presence of polysaccharides (85.00-181.00 mg g-1), water-soluble extractives (47.52-57.63%) and total ash (1.87-3.39%) in Radix Pseudostellariae. Polysaccharides and total ash showed no sensitivity to soil pH. Radix Pseudostellariae collected from soil with pH > 7 exhibited slightly higher levels of water-soluble extractives, Mg and Ca than that from soil with pH < 5. Conversely, soil with a pH less than 5 had higher OC, AN, AP and AK contents. Water-soluble extractives in Radix Pseudostellariae were negatively correlated with soil pH but positively correlated with OC and AN. The results imply that the sequestration of soil nutrients over long-term Pseudostellaria heterophylla cultivation could negatively impact the accumulation of some bioactive ingredients in Radix Pseudostellariae. This study has a profound implication for enhancing the quality of Radix Pseudostellariae of artificially cultivated Pseudostellaria heterophylla. © 2024 Society of Chemical Industry.

Structural characterisation of a novel polysaccharide from pseudostellaria heterophylla fibrous root and its anti-inflammatory and antioxidant activities in vitro

A novel water-soluble polysaccharide, named PF90-1, with a molecular weight of 1.8 kDa, was isolated and purified from the fibrous root of Pseudostellaria heterophylla. PF90-1 is composed of Gal, Glc and Man in a molar ratio of 73.61: 19.11: 7.28. Methylation analysis revealed that PF90-1 comprises of T-Galp, 1,4-Galp, 1,3,4-Galp, 1,2,3,4-Galp, T-Glcp and 1,3-Manp in a molar ratio of 37.89: 9.37: 17.01: 12.01: 15.88: 7.83. Bioactivity experiments showed that PF90-1 significantly improved lipopolysaccharide (LPS)-induced inflammatory damage in RAW264.7 cells by inhibiting nitric oxide (NO) production and reducing the levels of pro-inflammatory factors (IL-1β and TNF-α). In addition, PF90-1 exhibited strong antioxidant effects, protecting PC12 cells from H2O2-induced oxidative damage. This findings suggest that PF90-1 holds potential therapeutic value for the treatment of inflammatory and oxidative injuries.

Plant growth-promoting rhizobacteria enhance active ingredient accumulation in medicinal plants at elevated CO2 and are associated with indigenous microbiome.

Plant growth-promoting rhizobacteria (PGPR) and elevated CO2 (eCO2) have demonstrated their individual potential to enhance plant yield and quality through close interaction with rhizosphere microorganisms and plant growth. However, the efficacy of PGPR under eCO2 on rhizosphere microbiome and, ultimately, plant yield and active ingredient accumulation are not yet fully understood. This study investigated how the medicinal plant Pseudostellaria heterophylla (P. heterophylla) and its rhizosphere microbes respond to PGPR (Bacillus subtilis and Pseudomonas fluorescens) at eCO2 (1,000 ppm). It was found that the yield and active ingredient polysaccharides accumulation in the tuber of P. heterophylla were significantly increased by 38 and 253%, respectively. This promotion has been associated with increased root development and changes in the indigenous microbial community. Metagenomics analysis revealed a significant reduction in pathogenic Fusarium abundance in the rhizosphere. Potential biocontrol bacteria Actinobacteria and Proteobacteria were enriched, especially the genera Bradyrhizobium and Rhodanobacter. The reshaping of the rhizosphere microbiome was accompanied by the upregulation of biological pathways related to metabolite biosynthesis in the rhizosphere. These modifications were related to the promotion of the growth and productivity of P. heterophylla. Our findings highlighted the significant role played by PGPR in medicinal plant yield and active ingredient accumulation when exposed to eCO2.

Pathogen-driven Pseudomonas reshaped the phyllosphere microbiome in combination with Pseudostellaria heterophylla foliar disease resistance via the release of volatile organic compounds

BackgroundContinuous monocropping obstacles are common in plants, especially medicinal plants, resulting in disease outbreaks and productivity reductions. Foliar disease, mainly caused by Fusarium oxysporum, results in a severe decrease in the yield of Pseudostellaria heterophylla annually. Determining an effective biomethod to alleviate this disease is urgently needed to improve its productivity and quality.ResultsThis study screened thirty-two keystone bacterial genera induced by pathogens in P. heterophylla rhizosphere soil under continuous monocropping conditions. Pseudomonas, Chryseobacterium, and Flavobacterium, referred to as the beneficial microbiota, were significantly attracted by pathogen infection. The P. palleroniana strain B-BH16-1 can directly inhibit the growth and spore formation of seven primary pathogens of P. heterophylla foliar disease by disrupting fusaric acid production via the emission of volatile organic compounds (VOCs). In addition, strain B-BH16-1 enhances the disease resistance of P. heterophylla by obliterating the pathogen and assembling beneficial microbiota.ConclusionPathogen-induced Pseudomonas reshaped phyllosphere microbial communities via direct antagonism of pathogens and indirect disruption of the pathogen virulence factor biosynthesis to enhance disease suppression and improve yields. These results show that inhibiting pathogen virulence biosynthesis to reshape the plant microbial community using disease-induing probiotics will be an innovative strategy for managing plant disease, especially under continuous monoculture conditions.

Leaf grey spot caused by Arcopilus aureus on tobacco in China.

Tobacco (Nicotiana tabacum L.) is one of the most widely cultivated industrial crops worldwide. From April to July 2023, about 40% of tobacco seedlings in the greenhouse exhibited irregular taupe lesions in Zhengzhou, Henan Province, China. At an early stage of the lesion development, light grey spots with the diameter of 1-2 mm were observed, these spots gradually expanded and connected into large irregular lesions causing leaf wrinkling or withered. A total of 12 infected leaf tissues were sterilized with 75% ethanol for 45 s, rinsed three times in sterilized water and then plated on potato dextrose agar (PDA) medium for 10 days at 28°C in darkness. Seven fungal colonies that show the similar appearance were isolated and three of them (MB-1, MB-2 and MB-3) were used for subsequent identification. Colonies of these strains on PDA with loose mycelium and orange-red pigment on the underside, white aerial in the center and light yellow hyphae near the periphery, formed in the shape of a concentric ring pattern. Ascomata appeared from the 14th day, were black, spherical or ellipsoid with walls of textura angularis, and size was 53.8-101.1 μm × 50.3-104.3 μm (n=30). Terminal hairs were brown and straight, gradually tapering toward the tips. Asci clavate or fusiform, spore bearing part 16.2-29.2 × 7.3-11.4 μm (n=21), with 8 irregularly arranged ascospores, evanescent. Ascospores are brown at maturity, biapiculate, navicular or fusiform shapes with size of 8.7-12.8 μm × 4.8-6.9 μm (n=100), and more or less inaequilateral. Single spore strains derived from these strains exhibited the morphological features consistent with the original strains. The morphological characteristics of the fungus were consistent with the description of Arcopilus aureus (Chivers) X.W. Wang & Samson (= Chaetomium aureum Chivers) (Lee et al. 2019). Furthermore, the sequences of RPB2 region were amplified from these strains and the result sequences (GenBank accession no. OR513105-OR513108) all showed a 100.00% identity with A. aureus strain CBS 538.73 (GenBank accession no. KX976807.1). It was reported that the RPB2 gene was efficient in discriminating Arcopilus species (Tavares et al. 2022), thus a maximum likelihood (ML) phylogenetic tree based on the RPB2 gene sequences were constructed using MEGA 7.0 with 1000 replications of bootstrapping (Kumar et al. 2016), which revealed that these strains formed a well-supported clade with A. aureus strains of (CBS 153.52 and CBS538.73) (Wang et al. 2022). Pathogenicity analysis were performed on healthy flue-cured tobacco seedlings leaves (cv Y85) by using mycelial agar plugs (5 mm in diameter) and spore suspension (1×106 spores/mL), and the PDA plugs and sterile water were used for control group, respectively. Tobacco seedlings were incubated in a 25°C and 70% RH growth chamber. After seven days, the leaves showed obvious symptoms, with taupe lesions and yellow halos on the periphery, whereas no symptoms were found on the control leaves. The A. aureu was then reisolated from inoculated diseased leaves. Previously, A. aureus has been only reported to cause leaf black disease on Pseudostellaria heterophylla in China (Yuan et al. 2021). To our knowledge, this is the first reported of A. aureus causing tobacco leaf grey spot worldwide. Arcopilus aureus has been reported as a plant biocontrol fungus (Wang et al. 2013). However, due to the potential serious damage in tobacco seedlings caused by this fungus, the use of A. aureus as a plant biocontrol agent needs to be given more attention, and disease control measures of this pathogen should be developed.

Plant-soil hydraulic interaction and rhizosphere bacterial community under biochar and CO2 enrichment

The increasing atmospheric CO2 concentration is a global concern that affects the plant-bacteria-soil system. Previous studies have investigated plant growth and bacteria activity under CO2 enrichment. However, the effects of coupled elevated CO2 and biochar amendment on the interactions of soil and medicinal plants are not well understood. This study aims to investigate the medicinal plant-soil hydraulic interactions and rhizosphere bacteria communities under coupled CO2 enrichment and biochar conditions. Two levels of CO2 concentration (400, 1000 ppm) and two biochar dosages (3%, 5% by mass) were considered. Pseudostellaria heterophylla was used as the tested medicinal plant. During plant growth, coupled CO2 enrichment and biochar at 3% and 5% dosage increased the volumetric water content at a matric suction of 33 kPa by 97% and 82% respectively, which indicates enhanced water retention. The transpiration rate of P. heterophylla was slightly reduced by 11–30% with an increase in biochar dosage due to higher total suction, while it was significantly reduced by up to 57% due to CO2 enrichment. In the rhizosphere of P. heterophylla, elevated CO2 (1000 ppm) coupled with 3% biochar dramatically increase the relative abundance of Thaumarchaeota, which played an important role in C and N cycles. Moreover, coupled CO2 enrichment and biochar addition resulted in the highest bacterial richness, while 3% biochar at ambient CO2 induced the highest bacterial diversity. This study provides a basis for understanding the medicinal plant-bacteria-soil system under CO2 enrichment and biochar conditions.

Diversity of causal agents of Pseudostellaria heterophylla leaf spot from Guizhou Province, China

AbstractPseudostellaria heterophylla, a valuable Chinese herbal medicine, is widely cultivated in China, with a planted area of 5402.7 hm2 in Shibing County, Guizhou Province. The occurrence of leaf spot disease represents a significant challenge in the production area. In this study, 30 samples of P. heterophylla exhibiting symptoms of leaf spot were collected from three primary production areas in Guizhou Province. Following isolation and purification, 51 isolates were classified into two genera based on both cultural and conidial characteristics, along with ITS sequence data. Fourteen representative strains were selected for morphological and multigene molecular identification, which revealed that these representative strains were affiliated with Alternaria arborescens species complex (SC), Alternaria tenuissima, Alternaria gaisen, and Sclerotiophoma versabilis. The isolation rates of Alternaria and Sclerotiophoma were 84.3% and 15.7%, respectively. Pathogenicity tests confirmed the identity of the causal agents and demonstrated varying levels of aggressiveness among the different genera and species of pathogens towards P. heterophylla leaves. This study marks the first identification of A. arborescens SC, A. gaisen, and S. versabilis as pathogens causing leaf spot in P. heterophylla in China.

Pseudostellaria heterophylla improves intestinal microecology through modulating gut microbiota and metabolites in mice.

Pseudostellaria heterophylla is a Chinese medicine and healthy edible that is widely used to for its immunomodulatory, antioxidant, antidiabetic and antitussive properties. However, the potential function of P. heterophylla in intestinal microecology remains unclear. In this study, we investigated the impact of P. heterophylla on immune functions and evaluated its potential to regulate the gut microbiota and metabolome. The results showed that P. heterophylla significantly increased the content of red blood cells, total antioxidant capacity and expression of immune factors, and decreased platelet counts when compared to the control under cyclophosphamide injury. In addition, P. heterophylla altered the diversity and composition of the gut bacterial community; increased the abundance of potentially beneficial Akkermansia, Roseburia, unclassified Clostridiaceae, Mucispirillum, Anaeroplasma and Parabacteroides; and decreased the relative abundance of pathogenic Cupriavidus and Staphylococcus in healthy mice. Metabolomic analyses showed that P. heterophylla significantly increased the content of functional oligosaccharides, common oligosaccharides, vitamins and functional substances. Probiotics and pathogens were regulated by metabolites across 11 pathways in the bacterial-host co-metabolism network. We demonstrated that P. heterophylla increased the abundance of probiotics and decreased pathogens, and further stimulated host microbes to produce beneficial secondary metabolites for host health. Our studies highlight the role of P. heterophylla in gut health and provide new insights for the development of traditional Chinese medicine in the diet. © 2024 Society of Chemical Industry.

Synthesis of Pseudostellaria heterophylla polysaccharide-gold nanocomposites and their antitumor effect through immunomodulation.

Polysaccharides from natural sources have an excellent immune function and low toxicity; however, their limitations such as short half-life and instability limit their sustained pharmacological activity. In this context, the combination of polysaccharides and nanotechnology have been developed to promote the stability and prolong the immune activities of polysaccharides. To synthesize and explore the antitumor effect and immunomodulatory activity of PHP-AuNPs. Polysaccharides extracted from Pseudostellaria heterophylla were used to synthesize gold nanocomposites (PHP-AuNPs), and their physicochemical properties and immunoregulatory effect in vitro and in vivo were analyzed. The PHP-AuNPs were green synthesized with high biosafety. PHP-AuNPs can activate macrophages in vitro and decrease the tumor weight and volume, whereas they increase the immune organ index in vivo. Besides, PHP-AuNPs showed a beneficial effect for maintaining the immune balance of CD4+/CD8+ T cells and modulating the release of cytokines such as TNF-α increase and IL-10 decrease in mice. All these results suggested that PHP-AuNPs exhibit a remarkable antitumor effect and stronger immunomodulatory activity than that of free PHP-1. RESEARCH HIGHLIGHTS: The P. heterophylla polysaccharide-gold nanocomposites (PHP-AuNPs) were synthesized and physicochemical properties were characterized. The cytotoxicity in vitro and immunomodulatory effects of PHP-AuNPs on macrophages were analyzed. The immune-antitumor effects in vivo of PHP-AuNPs have also been confirmed.

First Report of Apiospora arundinis Causing Wet Root Rot of Pseudostellaria heterophylla in China.

Pseudostellaria heterophylla is one of the traditional medicines in China. From 2020 to 2022, postharvest wet root rot disease was observed with an incidence of 2~5% on the tuberous roots of the harvested P. heterophylla in Zherong county, Fujian province, China, which usually occurs under damp and unventilated conditions. The symptoms of the disease were as follows: white mycelia grew on the surface of tuberous root initially and gradually wrapped around the roots, the internal root tissue turned yellow and became wet decay finally. To identify the causal agent, a total of 20 samples with symptomatic tuberous roots were collected. Small pieces (3 mm2) were treated by surface disinfection with 75% ethanol and 1% NaOCl, then rinsed 3 times with sterile water. These treated pieces were transferred onto potato dextrose agar (PDA) and incubated at 25°C in the dark for 7 d. Ten pure cultures were obtained using single-spore isolation method. The fungus colonies initially produced white aerial mycelium, subsequently exhibited yellow pigmentation. Mycelia were consisted of smooth, hyaline, branched, and septate hyphae. The conidia were solitary or clustered, brown or dark brown, smooth, ellipsoidal to spherical, 6.66 (5.50-7.81)×5.65 (4.17-7.22) µm (n=50) in size. The conidiophores were hyaline or pale brown and produced conidiogenous cells, which were pale brown, smooth, ampulliform, and 10.14 (8.82-15.30) um long (n=50). Based on these morphological characteristics, the fungus was identified as the genus Apiospora (Arthrinium). The rDNA-ITS region and partial β-tubulin gene (BenA) were amplified using the primers ITS1/ITS4 (White et al. 1990) and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. The sequences of isolates FJAT-32563 and FJAT-32564 were deposited in GenBank (ITS, OM920984 and OM920985; BenA, OM953823 and OM953824). All sequences had more than 99% similarity with those of A. arundinis strain CBS:106.12 (ITS, KF144883; BenA, KF144973). In the multilocus phylogenetic analysis (ITS + BenA), the two isolates clustered together with other strains of A. arundinis with 100% bootstrap support. The isolates were therefore identified as A. arundinis based on both morphological and molecular characteristics. To confirm the pathogenicity, fresh tuberous roots were selected and surface disinfected, then the roots were immersed with a quarter length in the conidial suspension (106/mL) for 30 min, whereas the control roots were immersed with sterile water (n=30). They were placed in petri dish with wet filter paper at 25±2℃, maintaining 80% relative humidity in the dark. The white aerial mycelium appeared at 5 days after inoculation, and wet root rot decaying occurred after inoculation for 21 days. The symptoms were similar to those described above, whereas the control roots were asymptomatic. The same fungus was re-isolated from the infected roots, showing similar morphological characteristics and molecular traits. Koch's postulates were completed and the pathogenicity test for the isolates has been repeated thrice. Previously, A. arundinis was reported to infect peach and sugarcane (Ji et al. 2020; Liao et al. 2022). To our knowledge, this is the first report of A. arundinis causing wet root rot of P. heterophylla in China. The disease would be a potentially new threat to this medicinal plant.

Synergistic Effects of Oligochitosan and Pyraclostrobin in Controlling Leaf Spot Disease in Pseudostellaria heterophylla.

Pseudostellaria heterophylla (or Taizishen in Chinese), a medicinal, edible, and ornamental Chinese herb, is seriously affected by leaf spot disease (LSD). Oligochitosan is a natural agricultural antibiotic that is produced via the degradation of chitosan, which is deacetylated from chitin; pyraclostrobin is a broad-spectrum and efficient strobilurin fungicide. In this work, the ability of pyraclostrobin, oligochitosan, and their formula to manage P. heterophylla leaf spot disease and their role in its resistance, leaf photosynthesis, agronomic plant traits, root growth, and root quality were studied. The results show that the joint application of oligochitosan and low-dosage pyraclostrobin could control LSD more efficiently, with control effects of 85.75-87.49% compared to high-dosage pyraclostrobin or oligochitosan alone. Concurrently, the application of this formula could more effectively improve the resistance, leaf photosynthesis, agronomic plant traits, root yield, and medicinal quality of P. heterophylla, as well as reduce the application of pyraclostrobin. This finding suggests that 30% pyraclostrobin suspension concentrate (SC) 1500-time + 5% oligosaccharin aqueous solutions (AS) 500-time diluent can be recommended for use as a feasible formula to manage LSD and reduce the application of chemical pesticides.

Response of Soil Microorganisms and Phenolic to Pseudostelariae heterophylla Cultivation in Different Soil Types

Pseudostellaria heterophylla of the Caryophyllaceae family is an important medicinal herb in traditional Chinese medicine, but it encounters continuous cropping obstacle during cultivation. This study aimed to study the differences in the response of Pseudostellaria heterophylla cultivation to soil microorganisms and phenolic substances in different types of soil (Loamy soil and Clayey soil). The analysis of soil metabolites using HPLC-ESI-Q/TOF-MS and HPLC-DAD techniques revealed that the content of various phenolic components in the rhizosphere soil increased significantly after Pseudostellaria heterophylla cultivation. Pseudostellaria heterophylla cultivation in yellow soil could increase the abundance of Proteobacteria, Ascomycota, and Thermoplasmatota, while in brown soil, the abundance of Ascomycota significantly decreased. In addition, Pseudostellaria heterophylla cultivation could increase the α-diversity of bacteria and fungi in yellow soil, while reducing the α-diversity of archaea, and the impact on diversity in brown soil was relatively small. Correlation analysis showed that phenolic compounds were more likely to regulate the genera Bradyrhizobium, Calcarisporiella, Boothiomyces and Methanocella. Pseudostellaria heterophylla may regulate rhizosphere microbial diversity and community structure by secreting phenolic. However, in different types of soil environments, the response mechanism of soil microorganisms to Pseudostellaria heterophylla planting might vary. Our study provides new insights that the variability in the response of different soil types to Pseudostelariae heterophylla cultivation should be taken into account when developing strategies for reducing continuous cropping obstacles in medicinal plants.

Non-destructive identification of Pseudostellaria heterophylla from different geographical origins by Vis/NIR and SWIR hyperspectral imaging techniques.

The composition of Pseudostellaria heterophylla (Tai-Zi-Shen, TZS) is greatly influenced by the growing area of the plants, making it significant to distinguish the origins of TZS. However, traditional methods for TZS origin identification are time-consuming, laborious, and destructive. To address this, two or three TZS accessions were selected from four different regions of China, with each of these resources including distinct quality grades of TZS samples. The visible near-infrared (Vis/NIR) and short-wave infrared (SWIR) hyperspectral information from these samples were then collected. Fast and high-precision methods to identify the origins of TZS were developed by combining various preprocessing algorithms, feature band extraction algorithms (CARS and SPA), traditional two-stage machine learning classifiers (PLS-DA, SVM, and RF), and an end-to-end deep learning classifier (DCNN). Specifically, SWIR hyperspectral information outperformed Vis/NIR hyperspectral information in detecting geographic origins of TZS. The SPA algorithm proved particularly effective in extracting SWIR information that was highly correlated with the origins of TZS. The corresponding FD-SPA-SVM model reduced the number of bands by 77.2% and improved the model accuracy from 97.6% to 98.1% compared to the full-band FD-SVM model. Overall, two sets of fast and high-precision models, SWIR-FD-SPA-SVM and SWIR-FD-DCNN, were established, achieving accuracies of 98.1% and 98.7% respectively. This work provides a potentially efficient alternative for rapidly detecting the origins of TZS during actual production.

Integrated transcriptomic and proteomic analysis of exogenous abscisic acid regulation on tuberous root development in Pseudostellaria heterophylla.

Abscisic acid (ABA) significantly regulates plant growth and development, promoting tuberous root formation in various plants. However, the molecular mechanisms of ABA in the tuberous root development of Pseudostellaria heterophylla are not yet fully understood. This study utilized Illumina sequencing and de novo assembly strategies to obtain a reference transcriptome associated with ABA treatment. Subsequently, integrated transcriptomic and proteomic analyses were used to determine gene expression profiles in P. heterophylla tuberous roots. ABA treatment significantly increases the diameter and shortens the length of tuberous roots. Clustering analysis identified 2,256 differentially expressed genes and 679 differentially abundant proteins regulated by ABA. Gene co-expression and protein interaction networks revealed ABA positively induced 30 vital regulators. Furthermore, we identified and assigned putative functions to transcription factors (PhMYB10, PhbZIP2, PhbZIP, PhSBP) that mediate ABA signaling involved in the regulation of tuberous root development, including those related to cell wall metabolism, cell division, starch synthesis, hormone metabolism. Our findings provide valuable insights into the complex signaling networks of tuberous root development modulated by ABA. It provided potential targets for genetic manipulation to improve the yield and quality of P. heterophylla, which could significantly impact its cultivation and medicinal value.

Influences of phosphorus-modified biochar on bacterial community and diversity in rhizosphere soil.

Root-associated bacteria play a vital role in the soil ecosystem and plant productivity. Previous studies have reported the decline of bacterial community and rhizosphere soil quality in the cultivation of some medicinal plants (i.e., Pseudostellaria heterophylla). Phosphorus (P)-modified biochar has the potential to improve soil health and quality. However, its influence on the bacterial community and diversity in the rhizosphere of medicinal plants is not well understood. Therefore, this study aims to investigate the effects of P-modified biochar on the bacterial community and diversity in the rhizosphere of P. heterophylla. Soil samples were collected from the rhizosphere of 4-month P. heterophylla under control (no biochar), 3% unmodified and 3% P-modified biochar treatments, respectively. Compared with control and unmodified biochar treatment, P-modified biochar significantly increased the relative abundance of plant-beneficial bacteria (P < 0.05), particularly Firmicutes, Nitrospirae and Acidobacteria. The relative abundance of Bacillus, belonging to Firmicutes, was dramatically raised from 0.032% in control group to 1.723% in P-modified biochar-treated group (P < 0.05). These results indicate the potential enhancement of soil quality for the growth of medicinal plants. The application of biochar significantly increased bacterial richness and bacterial diversity (P < 0.05). P modification of biochar did not have significant effects on soil bacterial richness (P > 0.05), while it reduced Shannon and increased Simpson diversity index of soil bacterial communities significantly (P < 0.05). It indicates a decrease in bacterial diversity. This research provides a new perspective for understanding the role of P-modified biochar in the rhizosphere ecosystem.

Extraction of Polysaccharides from Root of Pseudostellaria heterophylla (Miq.) Pax. and the Effects of Ultrasound Treatment on Its Properties and Antioxidant and Immune Activities.

The hydrophilic polysaccharides (PS) were isolated and purified from the tuberous roots of Pseudostellaria heterophylla. The extraction process of PS from Pesudostellariae radix was optimized by single-factor experiments and orthogonal design. The extract was purified by DEAE cellulose column to obtain the pure polysaccharide PHP. Then PHP was treated with different intensities of sonication to study the effect of sonication on PHP's characteristics and its biological activity in vitro and in vivo. The results of this study revealed that ultrasound treatment did not significantly change the properties of PHP. Further, with the increase of ultrasound intensity, PHP enhanced the proliferation and phagocytosis of macrophage RAW264.7. Meanwhile, it could also significantly improve the body's antioxidant activity and immune function. The results of this study demonstrated that PHP has the potential as a food additive with enhanced antioxidant and immune functions, and its biological activities could be enhanced by sonication.

A Probiotic Bacillus amyloliquefaciens D-1 Strain Is Responsible for Zearalenone Detoxifying in Coix Semen.

Zearalenone (ZEN) is a mycotoxin produced by Fusarium spp., which commonly and severely contaminate food/feed. ZEN severely affects food/feed safety and reduces economic losses owing to its carcinogenicity, genotoxicity, reproductive toxicity, endocrine effects, and immunotoxicity. To explore efficient methods to detoxify ZEN, we identified and characterized an efficient ZEN-detoxifying microbiota from the culturable microbiome of Pseudostellaria heterophylla rhizosphere soil, designated Bacillus amyloliquefaciens D-1. Its highest ZEN degradation rate reached 96.13% under the optimal condition. And, D-1 can almost completely remove ZEN (90 μg·g-1) from coix semen in 24 h. Then, the D-1 strain can detoxify ZEN to ZEM, which is a new structural metabolite, through hydrolyzation and decarboxylation at the ester group in the lactone ring and amino acid esterification at C2 and C4 hydroxy. Notably, ZEM has reduced the impact on viability, and the damage of cell membrane and nucleus DNA and can significantly decrease the cell apoptosis in the HepG2 cell and TM4 cell. In addition, it was found that the D-1 strain has no adverse effect on the HepG2 and TM4 cells. Our findings can provide an efficient microbial resource and a reliable reference strategy for the biological detoxification of ZEN.

Dietary supplementation with pseudostellaria heterophylla polysaccharide enhanced immunity and changed mRNA expression of spleen in chicks

In recent years, increasing interest has focused on natural components extracted from plants, among which plant polysaccharides as natural immunomodulators that can promote animal immunity. The present study was performed to investigate the effect of feed supplement Pseudostellaria Heterophylla Polysaccharide (PHP) on serum Immunoglobulins, T lymphocyte subpopulations, Cytokines and Lysozyme (LZM) activity in chicks. In addition, the influence of PHP on splenic gene expression was investigated by transcriptome sequencing. Four hundred 7-day-old Gushi cocks were randomly divided into four groups in a completely randomized design. The chicks were fed with a basal diet supplemented with 0 (CON-A), 100 (PHP-L), 200 (PHP-M) and 400 (PHP–H) mg/kg PHP. Blood and spleen samples were collected from 6 randomly selected chicks in each group at 14, 21, 28, and 35 days of age. The results showed that compared to the CON-A group, the PHP-M group exhibited significant increases in the levels of IgA, IgG, IgM, CD3, and LZM in the serum at 14, 21, 28, and 35 days (P < 0.05), and at 28 d, there was a significant quadratic relationship between the levels of dietary PHP and the levels of IgG, IgM, IFN-γ, IL-2, CD3, and LZM. Furthermore, a total of 470 differentially expressed genes (DEGs) were identified in spleen from PHP-M and CON-A at 28 d. These DEGs were significantly enriched in the Phagosome, Intestinal immune network for IgA production and Cytokine-cytokine receptor interaction pathways. The present investigation highlights the ameliorating effect of dietary PHP on immunological variables and spleen of chicks, the study suggests that PHP supplementation can enhance immunity and positively impact spleen mRNA expression in chicks.

Quality control of Ganoderma lucidum by using C, H, O, and N stable isotopes and C and N contents for geographical traceability.

Ganoderma lucidum (G. lucidum) is a popular medicinal fungus that has been used in traditional medicine for decades, with its provenance influencing its medicinal and commercial worth. The amount of active ingredients and the price of G. lucidum from different origins vary significantly; hence, fraudulent labeling is common. Reliable techniques for G. lucidum geographic verification are urgently required to safeguard the interests of consumers, producers, and honest dealers. A stable isotope is widely acknowledged as a useful traceability technique and could be developed to confirm the geographical origin of G. lucidum. G. lucidum samples from various sources and in varying stages were identified by using δ 13C, δD, δ 18O, δ 15N, C, and N contents combined with chemometric tools. Chemometric approaches, including PCA, OPLS-DA, PLS, and FLDA models, were applied to the obtained data. The established models were used to trace the origin of G. lucidum from various sources or track various stages of G. lucidum. In the stage model, the δ 13C, δD, δ 18O, δ 15N, C, and N contents were considered meaningful variables to identify various stages of G. lucidum (bud development, growth, and maturing) using PCA and OPLS-DA and the findings were validated by the PLS model rather than by only four variables (δ 13C, δD, δ 18O, and δ 15N). In the origin model, only four variables, namely δ 13C, δD, δ 18O, and δ 15N, were used. PCA divided G. lucidum samples into four clusters: A (Zhejiang), B (Anhui), C (Jilin), and D (Fujian). The OPLS-DA model could be used to classify the origin of G. lucidum. The model was validated by other test samples (Pseudostellaria heterophylla), and the external test (G. lucidum) by PLS and FLDA models demonstrated external verification accuracy of up to 100%. C, H, O, and N stable isotopes and C and N contents combined with chemometric techniques demonstrated considerable potential in the geographic authentication of G. lucidum, providing a promising method to identify stages of G. lucidum.