Pseudostellaria Heterophylla Research Articles

De Novo Sequencing and Assembly Analysis of the Pseudostellaria heterophylla Transcriptome.

Pseudostellaria heterophylla (Miq.) Pax is a mild tonic herb widely cultivated in the Southern part of China. The tuberous roots of P. heterophylla accumulate high levels of secondary metabolism products of medicinal value such as saponins, flavonoids, and isoquinoline alkaloids. Despite numerous studies on the pharmacological importance and purification of these compounds in P. heterophylla, their biosynthesis is not well understood. In the present study, we used Illumina HiSeq 4000 sequencing platform to sequence the RNA from flowers, leaves, stem, root cortex and xylem tissues of P. heterophylla. We obtained 616,413,316 clean reads that we assembled into 127, 334 unique sequences with an N50 length of 951 bp. Among these unigenes, 53,184 unigenes (41.76%) were annotated in a public database and 39, 795 unigenes were assigned to 356 KEGG pathways; 23,714 unigenes (8.82%) had high homology with the genes from Beta vulgaris. We discovered 32, 095 DEGs in different tissues and performed GO and KEGG enrichment analysis. The most enriched KEGG pathway of secondary metabolism showed up-regulated expression in tuberous roots as compared with the ground parts of P. heterophylla. Moreover, we identified 72 candidate genes involved in triterpenoids saponins biosynthesis in P. heterophylla. The expression profiles of 11 candidate unigenes were analyzed by quantitative real-time PCR (RT-qPCR). Our study established a global transcriptome database of P. heterophylla for gene identification and regulation. We also identified the candidate unigenes involved in triterpenoids saponins biosynthesis. Our results provide an invaluable resource for the secondary metabolites and physiological processes in different tissues of P. heterophylla.

Interaction of Pseudostellaria heterophylla with Quorum Sensing and Quorum Quenching Bacteria Mediated by Root Exudates in a Consecutive Monoculture System.

Many plant-pathogenic bacteria are dependent on quorum sensing (QS) to evoke disease. In this study, the population of QS and quorum quenching (QQ) bacteria was analyzed in a consecutive monoculture system of Pseudostellaria heterophylla. The isolated QS strains were identified as Serratia marcescens with SwrIR-type QS system and exhibited a significant increase over the years of monoculture. Only one QQ strain was isolated from newly planted soil sample and was identified as Bacillus thuringiensis, which secreted lactonase to degrade QS signal molecules. Inoculation of S. marcescens to P. heterophylla root could rapidly cause wilt disease, which was alleviated by B. thuringiensis. Furthermore, the expression of lactonase encoded by the aiiA gene in S. marcescens resulted in reduction of its pathogenicity, implying that the toxic effect of S. marcescens on the seedlings was QS-regulated. Meanwhile, excess lactonase in S. marcescens led to reduction in antibacterial substances, exoenzymes, and swarming motility, which might contribute to pathogensis on the seedlings. Root exudates and root tuber extracts of P. heterophylla significantly promoted the growth of S. marcescens, whereas a slight increase of B. thuringiensis was observed in both samples. These results demonstrated that QS-regulated behaviors in S. marcescens mediated by root exudates played an important role in replanting diseases of P. heterophylla.

Structural Elucidation of a Novel Polysaccharide from Pseudostellaria heterophylla and Stimulating Glucose Uptake in Cells and Distributing in Rats by Oral.

The semi-refined polysaccharide of Pseudostellaria heterophylla is a complex polysaccharide that exhibits significantly hypoglycemic activities. A novel homogeneous polysaccharide, named as H-1-2, was isolated from the semi-refined polysaccharide. The mean molecular weight of H-1-2 was 1.4 × 104 Da and it was only composed of d-glucose monosaccharide. Structure elucidation indicated that H-1-2 contains pyranride, and has the characteristics of the α-iso-head configuration, a non-reducing end (T-), 4-, 1,6-, and 1,4,6-connection, in all four ways to connect glucose. H-1-2 was a type of glucan, where chemical combination exists in the main chain between 1→4 linked glucose, and contains a small amount of 1,6-linked glucose, which was in the branched chain. In vitro HepG2, 3T3-L1, and L6 cells were used to assess cellular glucose consumption and cellular glucose uptake by glucose oxidase, and the transport of 2-NBDG fluorescence probe results showed that H-1-2 could clearly increase glucose uptake and utilization in muscle and adipose cells, which is beneficial to screen for in the discovery of anti-diabetes lead compounds. H-1-2 was labeled with radioisotopes (99mTc-pertechnetate). 99mTc-labeled-H-1-2 was performed by SPECT/CT analysis images after oral administration in rats. At 4 h post ingestion, about 50% of the radioactivity was observed in the intestine. No significant radioactivity was found in the heart, liver, and kidney, conjecturing that absorption of 99mTc-labeled H-1-2 might, via intestinal mucosa, be absorbed into systemic circulation. This problem, as to whether the polysaccharide is absorbed orally, will need further examination.

Oligopeptides in plant medicines cited in Chinese Pharmacopoeia

In total, 23 plant plant medicined containing oligopeptides were cited in Chinese Pharmacopoeia (1 part) of 2015 version including Rubia cordifolia, Linum usitatissimum, Aster tataricus, Psammosilene tunicoides, Pseudostellaria heterophylla, Stellaria dichotoma, Vaccaria segetalis, Dianthus superbus, Celosia argentea, Lycii Cortex, Citrus medica, C. aurantium, Panax ginseng, Parmx notoginseng, Schisandra chinensis, Sparganium stoloniferum, Euryale ferox, Ophiopogon japonicas, Pinellia ternate, Achyranthes bidentata, Physalis alkekengi, Polygonatum odoratum, and Leonuri Fructus. There were 187 oligopeptides in plant medicines above as reported. Oligopeptides consisted mainly of linear peptides and cyclic peptides. The linear peptides included dipeptides, tripeptides and pentapeptides, and cyclic peptides included cyclic, bicyclic and tricyclic peptides. The number of residues of single cyclic peptides ranged from two to twelve. Bicyclic peptides were isolated mainly from R. cordifolia and C. argentea. Modern pharmacological study showed that oligopeptides had many pharmacological effects, including antitumor, anticoagulant, antibacterial, immune suppression and so on.

Abstract 2175: Anemarrhena rhizome and Schisandra chinensis inhibit viability and promote apoptosis in pancreatic cancer cell lines

Abstract Pancreatic cancer has one of the highest mortality rates of any cancer, with just a 6% 5-year survival rate. The current standard chemotherapy is gemcitabine, alone or in combination with 5-fluorouracil. However, the prognosis still remains poor due to apoptotic resistance, presenting a need for an adjuvant therapy option that will induce apoptosis within pancreatic tumor cells. Emerging evidence has supported the use of herbal extracts as alternative or adjuvant treatments for pancreatic cancer. In this study, we evaluated the effects on pancreatic cancer cells of a mix of 8 herbal extracts, selected for their high clinical usage and documented ability to alleviate pancreas-specific ailments such as type 2 diabetes, yet not examined for application in pancreatic cancer. The objective of this study was to determine the inhibitory effects of Astragalus membranaceus, Pseudostellaria heterophylla, Gypsum fibrosum, Anemarrhena rhizome, Solomon's seal rhizome, Rehmannia, Schisandra chinensis, and Cornus fruit extracts on pancreatic cancer cell viability in combination (referred to as 8-mix) and individually on pancreatic cancer cells. We also examined extracts previously demonstrated to inhibit pancreatic cancer cells, Cordyceps chinensis and American ginseng, as a control. The viability of pancreatic adenocarcinoma cell lines (BxPC-3 and Panc-1) and the human pancreatic islet: Panc-1 hybrid cell line (1.1B4) was evaluated in response to the 8-mix, C. chinensis, and A. membranaceus via MTT assay. To further characterize which herbs in the 8-mix were responsible for the reduction in cell viability, BxPC-3 and Panc-1 cells were assessed upon treatment with each individual herbal extract via MTT assay following a 12 hour exposure. To examine the impact of these herbs on a non-cancerous cell line, the T80 ovarian epithelial cell line was also evaluated. We measured the apoptotic activity of the 8-mix and two most potent individual herbs within the mix using a commercially available caspase 3/7 assay on BxPC-3 cells. Initial testing showed significant inhibitory effects in all pancreatic cell lines induced by the 8-mix in a dose-dependent manner. A. rhizome and S. chinensis were the most potent inhibitors of pancreatic cancer cell viability among the individual extracts. A. rhizome had the greatest apoptotic activity in the BxPC-3 cells compared to S. chinensis, the 8-mix, and vehicle-treated controls. Within the non-cancerous ovarian T80 cell line, A. rhizome and S. chinensis significantly inhibited cell viability. Taken together, our data suggests that A. rhizome induces apoptosis in pancreatic cancer cells and both A. rhizome and S. chinensis significantly inhibit viability in numerous pancreatic cancer cell lines. These herbs may therefore have potential as an adjuvant therapy in combination with traditional chemotherapeutic agents for the treatment of advanced pancreatic cancer. Citation Format: Catherine MarElia, Brant Burkhardt. Anemarrhena rhizome and Schisandra chinensis inhibit viability and promote apoptosis in pancreatic cancer cell lines. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2175.

Cloning and expression analysis of four carotenoid cleavage dioxygenases genes from Pseudostellaria heterophylla

To investigate the molecular mechanism of quality formation of Pseudostellaria heterophylla, the carotenoid cleavage dioxygenases (CCDs) genes were cloned from the transcriptome database of P. heterophylla, and analyzed them with bioinformatics analysis and expression analysis. The sequence length of four new gene were 1 617, 1 461, 1 746, 1 875 bp, and subsequently, named as PhCCD1,PhNCED2,PhNCED3 and PhCCD4 according to its genetic relationship with Arabidopsis thaliana. The sequence analysis showed that four new gene were all containing REP65 domains and binding sites of ferrous ion, such as histidine, glutamates and aspartates. Analysis phylogeny showed that PhNCED2 and PhNCED3 were the cluster of NCEDs, PhCCD1 and PhCCD4 were the cluster of CCDs. In addition, PhCCD1 and AtCDD1 of Arabidopsis thaliana, PhCCD4 and AtCCD4 of A. thaliana,PhNCED2, PhNCED3 and AtNCED3 of A. thaliana have high similarities. Analysis of real-time fluorescence quantitative showed that PhNCED2 and PhNCED3 were expressed mainly in underground part, the expression quantity of PhNCED2 reached the highest in fibrous root, PhNCED3 keeps higher in phloem and xylem, it may be the key enzymes of ABA biosynthesis genes. Moreover,PhCCD1 and PhCCD4 were expressed mainly in aerial part,the expression quantity of PhCCD1 reached the highest in leaf,PhCCD4 keeps higher in stem and leaf.It may be involved in the biosynthesis of carotenoids for P. heterophylla. The study obtained CDDs gene of P. heterophylla for the first time,this would lay the foundation of developing the response mechanism of P. heterophylla about external stress further,and then exploring the biological approach of quality formation in P. heterophylla.

Research on quality regionalization of cultivated Pseudostellaria heterophylla based on climate factors

Maxent model was applied in the study to filtering the climate factors layer by layer. Polysaccharides and pseudostellarin B the two internal quality evaluation index were combined to analyse the interlinkages between climate factors and chemical constituents in order to search for the critical climate factors of Pseudostellaria heterophylla. Then based on the key climate factors to explicit the quality spatial distribution of P. heterophylla. The results showed that polysaccharides and climatic factors had no significant correlation, suggesting that the indicator was not climate-driven metabolites. Pseudostellarin B could construct regression model with the precipitation. And quality regionalization results showed that pseudostellarin B content presented firstly increased and then decreased trend from southeast to northwest, which was the consistent change with precipitation. It clearly proposed that precipitation was the key climate factor, which affected the accumulation of cyclopeptide compound for Pseudostellariae Radix.

Breeding and extension of Pseudostellaria heterophylla new variety "Shitai No.1" in Guizhou province

Based on collections and researches of Pesudostellaria heterophylla germplasm resources from different areas of China, by using Shibing SB-4 provenance as materials, the new variety "Shitai No.1" was bred by mass selection, small plot variety comparative test, regional variety comparative test and field trial planting. Compared with "Qian taizishen No.1" and P. heterophylla land races. The disease and lodging resistance, root yield, polysaccharide content and the first grade rate of "Shitai No.1" have obvious advantages. In addition, it is relatively stable of yield in "Shitai No.1" in different places. It is demonstrated that "Shitai No.1" is a fine variety that adapt to the producing areas of P. heterophylla in Guizhou province, it is worthy to be promoted.

Quality evaluation and growing regionalization of Pseudostellaria heterophylla in Guizhou

This paper is aimed to study the potential ecological suitability regionalization of Pseudostellaria heterophylla for selecting GAP planting base location and designing rational production layout. The ecological factors and contribution rate were determined by using maximum entropy (Maxent) model. Then, the information entropy theory was used to determine the relative importance of each environmental factor, and thus to determine the most limiting habitat criteria. Finally, the probable spatial distribution of P. heterophylla was determined based on GIS spatial analysis of habitat conditions. Meanwhile, the optimal index range of ecological factors was quantified. The moderately and highly suitable habitats were mainly located in Shibing, Huangping, Cengong, the middle and east of Kaili, the south of Yuqing, the west of Tongren. The percentage of moderately and highly suitable habitats for P. heterophylla in the study area was 3.64%, and its area was 6 405.39 km². The results also showed that seven dominant ecological factors controlled the distribution of P. heterophylla. These factors included agrotype, the warmest rain, aspect, slope, the warmest and highest temperature, contents of soil organic carbon, and the driest month precipitation. The habitat suitability assessment model based on GIS and Maxent model theory could accurately evaluate the habitat suitability distribution of P. heterophylla in Guizhou. In addition, we recommended Cengong and Zhenyuan county in Guizhou province as the worthy developing potential planting areas.

Cloning and bioinformatics analysis of abscisic acid 8'-hydroxylase from Pseudostellariae Radix

Abscisic acid 8'-hydroxylase was one of key enzymes genes in the metabolism of abscisic acid (ABA). Seven menbers of abscisic acid 8'-hydroxylase were identified from Pseudostellaria heterophylla transcriptome sequencing results by using sequence homology. The expression profiles of these genes were analyzed by transcriptome data. The coding sequence of ABA8ox1 was cloned and analyzed by informational technology. The full-length cDNA of ABA8ox1 was 1 401 bp,with 480 encoded amino acids. The predicated isoelectric point (pI) and relative molecular mass (MW) were 8.55 and 53 kDa,respectively. Transmembrane structure analysis showed that there were 21 amino acids in-side and 445 amino acids out-side. High level of transcripts can detect in bark of root and fibrous root. Multi-alignment and phylogenetic analysis both show that ABA8ox1 had a high similarity with the CYP707As from other plants,especially with AtCYP707A1 and AtCYP707A3 in Arabidopsis thaliana. These results lay a foundation for molecular mechanism of tuberous root expanding and response to adversity stress.

Insight into structure dynamics of soil microbiota mediated by the richness of replanted Pseudostellaria heterophylla.

Consecutive monoculture of crops causes serious diseases and significant decline in yield and quality, and microbes in the rhizosphere are closely linked with plant health. Here we systematically studied the structure dynamics of soil microbiota in the monocropping system of Pseudostellaria heterophlla. The results illustrated that the successive cropping of P. heterophylla shifts the diversity and structure of microbial community in rhizosphere soil of P. heterophylla, showing that the diversity of microbial community in rhizosphere soil of P. heterophylla was decreased with the increase of planting years while the structure of microbial community became more deteriorative. Moreover, the population size of typical pathogens increased and the beneficial bacterial population decreased with the increasing years of monoculture, which resulted in the microecological imbalance in P. heterophylla rhizosphere, thereby caused serious replanting diseases in monocropping system. Our results suggested that structure dynamics of rhizosphere microbial communities were mediated by the richness of replanted P. heterophylla, and thus the replant disease result from the imbalanced microbial structure with a higher ratio of pathogens/beneficial bacteria in rhizosphere soil under monocropping regimes. This finding provides a clue to open a new avenue for modulating the root microbiome to enhance the crop production and sustainability.

Effects of consecutive monoculture of Pseudostellaria heterophylla on soil fungal community as determined by pyrosequencing.

Under consecutive monoculture, the biomass and quality of Pseudostellaria heterophylla declines significantly. In this study, a three-year field experiment was conducted to identify typical growth inhibition effects caused by extended monoculturing of P. heterophylla. Deep pyrosequencing was used to examine changes in the structure and composition of soil fungal community along a three-year gradient of monoculture. The results revealed a distinct separation between the newly planted plot and the two-year, three-year monocultured plots. The Shannon and Simpson diversity indices were significantly higher in the two-year and three-year monoculture soils than in the newly planted soil. Consecutive monoculture of this plant led to a significant increase in relative abundance of Fusarium, Trichocladium and Myrothecium and Simplicillium, etc., but a significant decrease in the relative abundance of Penicillium. Quantitative PCR analysis confirmed a significant increase in Fusarium oxysporum, an agent known to cause wilt and rot disease of P. heterophylla. Furthermore, phenolic acid mixture at a ratio similar to that found in the rhizosphere could promote mycelial growth of pathogenic F. oxysporum. Overall, this study demonstrated that consecutive monoculture of P. heterophylla can alter the fungal community in the rhizosphere, including enrichment of host-specific pathogenic fungi at the expense of plant-beneficial fungi.

ITRAQ-based quantitative proteomic analysis of cultivated Pseudostellaria heterophylla and its wild-type

Pseudostellaria heterophylla is an important traditional Chinese herbal medicine with vast clinical consumption because of its positive effects. To date, changes in the metabolite composition of P. heterophylla have been well documented; however, the molecular differences between cultivated P. heterophylla and its wild-type are still unknown. The aim of this study was to investigate differences in cultivated and wild P. heterophylla. Due to the lack of a genomic database, we used high-throughput transcriptomic and proteomic technologies to identify proteins in the herb. Isobaric tags for relative and absolute quantification (iTRAQ) MS/MS were used to detect statistically significant changes between cultivated and wild P. heterophylla. We detected 3775 proteins; 332 showed differential accumulations across two different ecotypes of P. heterophylla. 71 significant differential expressions of proteins were selected based on GO annotations, KEGG, STRING analysis, and expression level. Quantitative real-time PCR analysis confirmed the authenticity and accuracy of the proteomic analysis. The results indicated that the carbohydrate and cellular amino acid metabolisms in cultivated P. heterophylla were weaker than those in its wild-type; seven important proteins were found to regulate sucrose and amino acids. This will provide the basic information for exploring the cause of differences in secondary metabolites in different ecotypes of P. heterophylla and the protein mechanism of its quality formation. Biological significanceThis study combined a transcriptome, proteome, and metabolism approach for analyzing differentially expressed proteins of cultivated and wild P. heterophylla and established the relationship between significantly differentially expressed proteins and differential chemical components in non-model plants. The results of proteomic analysis provide the basic information for exploring the quality forming process, which will demonstrate, and provide guidance for, the study of effective constituents of P. heterophylla and its quality formation mechanism.

Analysis of heavy metals in Pseudostellaria heterophylla in Baiyi Country of Wudang District

To understand the correlation between the contents of five heavy metals (including arsenic, As; cadmium, Cd; copper, Cu; mercury, Hg; and lead, Pb) and Pseudostellaria heterophylla quality in Baiyi Country of Wudang District, nine samples of P. heterophylla and planting soil were collected in the research area. After pre-treatment, the samples were sent to ALS Minerals–ALS Chemex (Guangzhou) Co. Ltd., for inductively coupled plasma atomic emission spectrometry (ICP-AES) and inductively coupled plasma mass spectrometry (ICP-MS) to determine their heavy metal contents. The average contents of the heavy metals in P. heterophylla are not higher than the limited standard value (LSV) of the heavy metals, and they generally conform to quality requirements. Except for Cd, the bioconcentration factors (BFs) of the heavy metals are not higher than 0.5, and those for Pb and As are especially low. Cd is slightly enriched in P. heterophylla and the other heavy metals are not. There is a significant positive correlation between the Cu and As in planting soil, a more significant positive correlation among of As, Cd and Pb in P. heterophylla, and the correlations among the other elements are not significant. The levels of enrichment of As, Cd, Cu and Pb contents are very severe, minor, not enriched and moderate in planting soil, respectively. As and Pb are mainly derived from anthropogenic activities. In particular, very severe As pollution by anthropogenic sources occurs. In addition, Cd and Cu are mainly from naturally occurring sources, and Cd may be impacted by anthropogenic activities.

A split G-quadruplex DNAzyme based magnetic graphene oxide platform for sensitive authentication of Pseudostellaria heterophylla

Based on the different affinities of graphene oxide (GO) toward ssDNA and dsDNA, a sensitive fluorescence assay utilizing a split G-rich probe and magnetic GO (Fe3O4/GO) was developed for authentication of Pseudostellaria heterophylla based on the ITS sequences.

HPLC-ESI-MS(n) Analysis, Fed-Batch Cultivation Enhances Bioactive Compound Biosynthesis and Immune-Regulative Effect of Adventitious Roots in Pseudostellaria heterophylla.

A electrospray ionization tandem mass spectrometry (ESI-MS(n)) analysis was performed in order to identify the active composition in Pseudostellaria heterophylla adventitious roots. Pseudostellarin A, C, D, and G were identified from P. heterophylla adventitious roots on the basis of LC-MS(n) analysis. The culture conditions of adventitious roots were optimized, and datasets were subjected to a partial least squares discriminant analysis (PLS-DA), in which the growth ratio and some compounds showed a positive correlation with an aeration volume of 0.3 vvm and inoculum density of 0.15 %. Fed-batch cultivation enhanced the contents of total saponin, polysaccharides, and specific oxygen uptaker rate (SOUR). The maximum dry root weight (4.728 g l(-1)) was achieved in the 3/4 Murashige and Skoog (MS) medium group. PLS-DA showed that polysaccharides contributed significantly to the clustering of different groups and showed a positive correlation in the MS medium group. The delayed-type hypersensitivity (DTH) reaction on the mice induced by 2,4-dinitrofluorobenzene (DNFB) was applied to compare the immunocompetence effects of adventitious roots (AR) with field native roots (NR) of P. heterophylla. As a result, AR possessed a similar immunoregulation function as NR.

LC-ESI-MS/MS analysis and pharmacokinetics of heterophyllin B, a cyclic octapeptide from Pseudostellaria heterophylla in rat plasma.

Heterophyllin B (HB) is a cyclic octapeptide isolated from Pseudostellaria heterophylla. HB is used as the quality control index for evaluating P. heterophylla in the Chinese Pharmacopoeia. A rapid and sensitive LC-ESI-MS/MS method was developed and validated for the analysis of HB in rat plasma. Sample preparation consisted of a solid-phase extraction step for the removal of interference and preconcentration of the target analyte HB and the internal standard N-acetylcysteine before chromatographic analysis by MS/MS detection. The separation of HB and N-acetylcysteine was performed using a Hypersil GOLD™ C18 column and a mixture of methanol-water (60:40, v/v) containing 10 mmol/L ammonium formate and 0.1% formic acid as the mobile phase. The determination step was optimized in the selected reaction monitoring mode for the highly selective and sensitive quantitation of HB in rat plasma. Intra- and inter-assay precision (as relative standard deviation) was ≤9.1%, and accuracy was between 92.6 and 102.7%. The validated method was successfully applied to quantify HB concentrations up to 7 h after tail intravenous injections of 2.08, 4.16 and 8.32 mg/kg HB in rats. The LC-MS/MS method identified the relevant pharmacokinetic parameters of HB and its studied analog.

Interaction of Pseudostellaria heterophylla with Fusarium oxysporum f.sp. heterophylla mediated by its root exudates in a consecutive monoculture system.

In this study, quantitative real-time PCR (qPCR) was used to determine the amount of Fusarium oxysporum, an important replant disease pathogen in Pseudostellaria heterophylla rhizospheric soil. Moreover, HPLC was used to identify phenolic acids in root exudates then it was further to explore the effects of the phenolic acid allelochemicals on the growth of F. oxysporum f.sp. heterophylla. The amount of F. oxysporum increased significantly in P. heterophylla rhizosphere soil under a consecutive replant system as monitored through qPCR analysis. Furthermore, the growth of F. oxysporum f.sp. heterophylla mycelium was enhanced by root exudates with a maximum increase of 23.8%. In addition, the number of spores increased to a maximum of 12.5-fold. Some phenolic acids promoted the growth of F. oxysporum f.sp. heterophylla mycelium and spore production. Our study revealed that phenolic acids in the root secretion of P. heterophylla increased long with its development, which was closely related to changes in rhizospheric microorganisms. The population of pathogenic microorganisms such as F. oxysporum in the rhizosphere soil of P. heterophylla also sharply increased. Our results on plant-microbe communication will help to better clarify the cause of problems associated with P. heterophylla under consecutive monoculture treatment.

A split G-quadruplex and graphene oxide-based low-background platform for fluorescence authentication of Pseudostellaria heterophylla.

A label-free split G-quadruplex and graphene oxide (GO)-based fluorescence platform has been designed to distinguish Pseudostellaria heterophylla (PH) from its adulterants based on the differences in their nrDNA ITS sequences. Herein, GO has been first introduced to capture G-rich probes with 2:2 split mode and then decrease the background signal. As T-DNA exists, the probes leave the GO surface to form double-stranded structures followed by the formation of the overhanging G-rich sequence into a G-quadruplex structure, which combines quinaldine red specifically to produce a strong fluorescence signal. In addition, this strategy allows detection of T-DNA in a wide range of concentrations from 1.0 × 10−8 to 2.0 × 10−6 mol·L−1 with a detection limit of 7.8 × 10−9 mol·L−1. We hope that the split G-quadruplex/GO platform can be utilized to further develop gene identification sensors in Traditional Chinese Medicine or other analysis areas.

Breeding on eight strains of Pseudostellaria heterophylla based on phenotypic traits and quality in Guizhou province

To provide new germplasm materials for breeding new varieties of Pseudostellaria heterophylla. The method of single plant selection was adopted, with the comparative experiments being carried out under the same conditions in Shibing county. The 8 plants of Shibing SB-4 were compared respectively with factor analysis for 27 phenotypic traits and 8 yield traits, and single factor variance analysis for the contents of polysaccharides. Using factor analysis, 27 phenotypic traits were classified into 7 principal divisors and 8 yield traits were simplified into 3 principal divisors. The 4 strains of P. heterophylla, ZT-01, ZT-02, ZT-06 and ZT-07, performed better than others in the phenotypic traits, and ZT-01, ZT-02, ZT-03 and ZT-07 in the yield traits. The contents of polysaccharides of ZT-01, ZT-02, ZT-05 and ZT-08 showed significantly higher value. There is significant difference among the 8 strains of P. heterophylla in phenotypic traits, yield traits and quality traits, making it possible to select certain strains for different purposes. ZT-01 and ZT-02 can be breaded further. ZT-06 and ZT-07 were used as ornamental cultivars for its great phenotypic traits. ZT-03 with good resistance and high yield was taken as resistant variety, and ZT-05 would face next selection on the basis of its high content of polysaccharide.