Screening of cadmium resistant bacteria and their growth promotion of Sorghum bicolor (L.) Moench under cadmium stress.

Heavy metal pollution of agricultural soils, especially from cadmium (Cd) contaminationcaused serious problems in both food security and economy. Sorghum bicolor (L.) showed a great potential in phytoremediation of Cd contamination due to its fast growth, high yield and easy harvesting. However, the growth of S. bicolor plants tends to be inhibited under Cd exposure, which limited its application for Cd remediation. Plant growth-promoting rhizobacteria may enhance the Cd resistance of S. bicolor and thus improve its Cd removal efficiency. In this study, three Cd-resistant bacteria were screened based on Cd and acid tolerance and identified as Bacillus velezensis QZG6, Enterobacter cloacae QZS3 and Bacillus cereus QZS8, by 16S rRNA sequencing. Inoculation of hydroponic plants with strains QZG6, QZS3 or QZS8 significantly promoted the biomass of sorghum plants by 31.52%, 50.20% and 26.93%, respectively, compared with those of uninoculated plants under Cd exposure. The activity of SOD, POD and MDA content in Cd-stressed S. bicolor plants were reduced of 65.74%, 31.52%, and 80.91%, respectively, when inoculated with the strains QZS3. For pot experiment, strains QZG6, QZS3 and QZS8 significantly promoted the biomass of sorghum plants by 47.30%, 19.27% and 58.47%, compared with those of uninoculated plants under Cd exposure. The activity of SOD, POD and MDA content in Cd-stressed S. bicolor plants were reduced of 67.20%, 22.40%, and 40.65%, respectively, when inoculated with the strains QZS3. All these three strains significantly increased the Cd removal efficiency of the plants by 42.16% (QZG6), 18.76% (QZS3) and 21.06% (QZS8). To investigate the bacterial characteristics associated with growth promotion of S. bicolor plants, the ability on nitrogen fixation, phosphorus solubilization, siderophores production, and phytohormones production were determined. All the strains were able to fix nitrogen. Phosphorus release was observed for strains QZG6 (inorganic or organic phosphorus) and QZS3 (inorganic phosphorus). Both QZG6 and QZS8 were able to produce siderophores, while only QZG6 was positive for ACC deaminase. All the strains produced IAA, SA and GA. These results indicated that the three strains promoted the plant growth under Cd stress, probably through Cd detoxification by siderophores, as well as through growth regulation by N/P nutrient supply and phytohormone. The present study showed a great potential of the three Cd-resistant strains combined with S. bicolor plants in the remediation of Cd-polluted soils, which may provide a new insight into combining the advantages of microbes and plants to improve the remediation of Cd-contaminated soils.

Biomechanical Evaluation of an Adaptive-Motion Pedicle Screw Fixation System: Experimental and Numerical Analysis.

Rigid fixation is mostly used in thoracolumbar spine surgery, which restricts the thoracolumbar spine segments moving and is not conducive to postoperative rehabilitation. We developed an adaptive-motion pedicle screw and established a finite element model of the T12-L3 segments of the thoracolumbar spine in osteoporosis patients based on the CT image data. A variety of internal fixation finite element models were established for mechanical simulation analysis and comparison. The simulation results showed that compared with the conventional internal fixation system, the mobility of the new adaptive-motion internal fixation system was improved by about 13.8% and 7.7% under the classic conditions such as lateral bending and flexion. in vitro experiments were conducted simultaneously with fresh porcine thoracolumbar spine vertebrae, and the axial rotation condition was taken as an example to analyze the mobility. The in vitro results showed that the mobility of the adaptive-motion internal fixation system had better mobility characteristics under axial rotation conditions, which was consistent with the finite element analysis. The adaptive-motion pedicle screws can preserve a certain degree of vertebral mobility, and avoid excessive vertebral restriction. It also increases the stress value of the intervertebral disk, which is closer to the normal mechanical transmission of the human body, avoiding stress masking and slowing down the degeneration of the intervertebral disk. The adaptive-motion pedicle screws can reduce the peak stress of the implant and avoid surgical failure due to implant fracture.

Physiological and transcriptome analysis of Dendrobium officinale under low nitrogen stress.

Nitrogen (N) is the main nutrient of plants, and low nitrogen usually affects plant growth and crop yield. The traditional Chinese herbal medicine Dendrobium officinale Kimura et. Migo is a typical low nitrogen-tolerant plant, and its mechanism in response to low nitrogen stress has not previously been reported. In this study, physiological measurements and RNA-Seq analysis were used to analyse the physiological changes and molecular responses of D. officinale under different nitrogen concentrations. The results showed that under low nitrogen levels, the growth, photosynthesis and superoxide dismutase activity were found to be significantly inhibited, while the activities of peroxidase and catalase, the content of polysaccharides and flavonoids significantly increased. Differentially expressed genes (DEGs) analysis showed that nitrogen and carbon metabolisms, transcriptional regulation, antioxidative stress, secondary metabolite synthesis and signal transduction all made a big difference in low nitrogen stress. Therefore, copious polysaccharide accumulation, efficient assimilation and recycling of nitrogen, as well as rich antioxidant components play critical roles. This study is helpful for understanding the response mechanism of D. officinale to low nitrogen levels, which might provide good guidance for practical production of high quality D. officinale .

Complete genome of Sphingomonas paucimobilis ZJSH1, an endophytic bacterium from Dendrobium officinale with stress resistance and growth promotion potential.

Sphingomonas paucimobilis ZJSH1 is an endophytic bacterium isolated from the roots of Dendrobium officinale with the ability to promote plant growth. It was found that the genome of strain ZJSH1 had gene fragment rearrangement compared with the genomes of the other four strains of S. paucimobilis, and the genome was integrated with phage genes. Functional analysis showed that the strain contained colonization-related genes, chemotaxis and invasion. A variety of genes encoding active materials, such as hormones (IAA, SA, ABA and zeaxanthin), phosphate cycle, antioxidant enzymes, and polysaccharides were identified which provide the strain with growth promotion and stress-resistant characteristics. Experiments proved that S. paucimobilis ZJSH1 grew well in media containing 80 g/L sodium chloride, 240 g/L polyethylene glycol and 800 µmol/L Cd2+, indicating its potential for resistance to stresses of salt, drought and cadmium, respectively. S. paucimobilis ZJSH1 is the only endophytic bacterium of this species that has been reported to promote plant growth. The analysis of its genome is conducive to understanding its growth-promoting mechanism and laying a foundation for the development and utilization of this species in the field of agriculture.

Differential Expression of Genes Related to Growth and Aflatoxin Synthesis in Aspergillus flavus When Inhibited by Bacillus velezensis Strain B2.

Aspergillus flavus is a saprophytic soil fungus that infects and contaminates seed crops with the highly carcinogenic aflatoxin, which brings health hazards to animals and humans. In this study, bacterial strains B1 and B2 isolated from the rhizosphere soil of camellia sinensis had significant antagonistic activities against A. flavus. Based on the phylogenetic analysis of 16SrDNA gene sequence, bacterial strains B1 and B2 were identified as Bacillus tequilensis and Bacillus velezensis, respectively. In addition, the transcriptome analysis showed that some genes related to A. flavus growth and aflatoxin synthesis were differential expressed and 16 genes in the aflatoxin synthesis gene cluster showed down-regulation trends when inhibited by Bacillus velezensis strain B2. We guessed that the Bacillus velezensis strain B2 may secrete some secondary metabolites, which regulate the related gene transcription of A. flavus to inhibit growth and aflatoxin production. In summary, this work provided the foundation for the more effective biocontrol of A. flavus infection and aflatoxin contamination by the determination of differential expression of genes related to growth and aflatoxin synthesis in A. flavus when inhibited by B. velezensis strain B2.

Paracandidimonas lactea sp. nov., a urea-utilizing bacterium isolated from landfill.

A urea-utilizing bacterium, designated Q2-2 T, was isolated from landfill. Cells of strain Q2-2 T were Gram stain-negative, aerobic, short-rod bacteria. Strain Q2-2 T was observed to grow at a temperature range of 15-37℃ (optimum 30 â„ƒ), a pH range of 5.5-9.5 (optimum pH 8.0) and 0-4% (w/v) NaCl (optimum 1%). The major respiratory quinone was Q-8, and the major polar lipids were diphosphatidyl glycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, and phosphatidyl glycerol. Based on the 16S rRNA gene sequence, strain Q2-2 T had the highest similarity with Paracandidimonas caeni 24 T (98.0%), followed by Pusillimonas soli MJ07T (97.5%), Parapusillimonas granuli Ch07T (97.2%), Pusillimonas ginsengisoli DCY25T (97.1%) and Paracandidimonas soli IMT-305 T (96.4%). The ANI values between strain Q2-2 T and the above related type strains were 71.02%, 73.52%, 74.32%, 74.59% and 72.29%, respectively. The DNA G + C content of strain Q2-2 T was 61.1%. Therefore, strain Q2-2 T represents a novel species of the genus Paracandidimonas, for which the name Paracandidimonas lactea sp. nov. (type strain Q2-2 T = CGMCC 1.19179 T = JCM 34906 T) is proposed.

Directional bioconversion and optimization of stevioside into rubusoside by Lelliottia sp. LST-1.

AIMS: The present study aimed to specifically transform stevioside (ST) into rubusoside (RS) through bioconversion with high efficiency, seeking to endow steviol glycosides (SGs) with subtle flavours for commercial acceptability. METHODS AND RESULTS: An endophytic bacterium named Lelliottia LST-1 was screened and confirmed to specifically convert ST into RS, reaching a conversion rate of 75.4% after response surface optimization. Phylogenetic analysis combined with complete genome sequencing demonstrated that LST-1 was also presumed to be a new species. To further explore the principle and process of biological transformation, the potential beta-glucosidases GH3-1, GH3-2, GH3-3 and GH3-4 were expressed, purified and reacted with SGs. High-performance liquid chromatography revealed that all enzymes hydrolysed ST and generated RS, but substrate specificity analysis indicated that GH3-2 had the highest substrate specificity towards STs and the highest enzyme activity. CONCLUSION: The potential ß-glucosidase GH3-2 in Lelliottia sp. LST-1 was found to specifically and efficiently convert ST to RS. SIGNIFICANCE AND IMPACT OF STUDY: The efficient biotransformation of ST into RS will be beneficial to its large-scale production and extensive application in the food and pharmaceutical industries.

Antagonistic and plant growth promotion effects of Mucor moelleri, a potential biocontrol agent.

With the increasing demand for high quality and environmentally safe or green food, Biological Control Agents (BCAs) are playing critical roles in green agriculture, which in turn has paved the way for the requirement of effective, appropriate microbial antagonists. In this study, Mucor moelleri AA1 was isolated and investigated for its growth promotion and antagonism against Athelia rolfsii and Colletotrichum gloeosporiodes. The results showed a high antagonistic activity of M. moelleri against A. rolfsii and C. gloeosporiodes with percentage inhibitions of 73 % and 86 % respectively using the dual plate method, and the same antagonistic activity was also observed in liquid cocultures. A pot study analysis showed significant suppression of the diseases as well as growth promotion on tomato. Scanning electron microscopy (SEM) indicated that M. moelleri inhibited the growth of mycelium and the production of web-like materials. Based on headspace-solid phase microextraction (HS-SPME) analysis, microbial volatile compounds were determined, which were mainly aromatic compounds and alkaloids. Also, several antagonistic enzymes, such as ß-1, 3- glucanase, proteases, catalase and ACC deaminase as well as the phytohormone IAA, were found to be produced by M. moelleri. Overall, these results combine to make M. moelleri a good prospective candidate for biological control and as a plant growth-promoting agent. The present study appears to be the first report identifying M. moelleri as a biological control agent.

Analysis of saponins detoxification genes in Ilyonectria mors-panacis G3B inducing root rot of Panax notoginseng by RNA-Seq.

Saponins are kinds of antifungal compounds produced by Panax notoginseng to resist invasion by pathogens. Ilyonectria mors-panacis G3B was the dominant pathogen inducing root rot of P. notoginseng, and the abilities to detoxify saponins were the key to infect P. notoginseng successfully. To research the molecular mechanisms of detoxifying saponins in I. mors-panacis G3B, we used high-throughput RNA-Seq to identify 557 and 1519 differential expression genes (DEGs) in I. mors-panacis G3B with saponins treatments for 4H (Hours) and 12H (Hours) compared with no saponins treatments, respectively. Among these DEGs, we found 93 genes which were simultaneously highly expressed in I. mors-panacis G3B with saponins treatments for 4H and 12H, they mainly belong to genes encoding transporters, glycoside hydrolases, oxidation-reduction enzymes, transcription factors and so on. In addition, there were 21 putative PHI (Pathogen-Host Interaction) genes out of those 93 up-regulated genes. In this report, we analyzed virulence-associated genes in I. mors-panacis G3B which may be related to detoxifying saponins to infect P. notoginseng successfully. They provided an excellent starting point for in-depth study on pathogenicity of I. mors-panacis G3B and developed appropriate root rot disease management strategies in the future.

Foetidibacter luteolus gen. nov., sp. nov., isolated from the rhizosphere of Salvia miltiorrhiza.

A polyphosphate-producing bacterium, YG09T, was isolated from the rhizosphere of Salvia miltiorrhiza. Its colonies were 2.0-3.0 mm in diameter, smooth, circular, convex and yellow after growth on R2A at 28 °C for 72 h, with aerobic, Gram-stain-negative, non-motile and rod-shaped bacteria. The strain was found to grow at 10-40 °C (optimum 37 °C), pH 5.5-8.0 (optimum 6.0), with 0-0.6% (w/v) NaCl (optimum 0). Chemotaxonomic analysis showed menaquinone-7 as the only quinone present; C15: 1 iso G, C15: 1 iso, C16: 0, C16: 0 3OH, C17: 0 iso 3OH, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) as the major fatty acids (> 5%), and phosphatidylethanolamine, three unidentified phospholipids, four unidentified polar lipids, three unidentified aminolipids, and one unidentified amino phospholipid as the polar lipids. The DNA G + C content was 44.6 mol%. The 16S rRNA gene sequences of the isolate showed highest similarities to Panacibacter ginsenosidivorans Gsoil 1550T (93.6%), Filimonas endophytica SR2-06T (93.4%), Parasegetibacter terrae SGM2-10T (92.8%), and Arvibacter flaviflagrans C-1-16T (92.7%), within the family Chitinophagaceae of the phylum Bacteroidetes. The ANI values between strain YG09T and Panacibacter ginsenosidivoran Gsoil 1550T, Filimonas endophytica SR2-06T and Filimonas lacunae YT21T were 69.4, 68.3 and 68.7%, respectively. Based on phenotypic, genotypic and phylogenetic analyses, strain YG09T represents a novel genus in the family Chitinophagaceae, for which the name Foetidibacter luteolus gen. sp. nov. is proposed. The type strain is Foetidibacter luteolus YG09T (= MCCC 1K04042T = KCTC 72595T).

MiR-381-3p/RAB2A axis activates cell proliferation and inhibits cell apoptosis in bladder cancer.

Acting as a really common cancer in the world, bladder cancer has taken many people's life away. MiRNAs and mRNA have been reported can regulate the expression of cancers. In this study, the role of RAB2A and miR-381-3p was fully studied in bladder cancer. qRT-PCR assay probe the expression of RAB2A and miR-381-3p in bladder cancer cells. Meanwhile, colony formation assay, EdU assay, flow cytometry analysis, JC-1 assay and western blot assay were implemented to detect the progression of bladder cancer cells. Silenced RAB2A could reduce the cell proliferation of bladder cancer, and activate the apoptosis. Meanwhile, miR-381-3p could bind to RAB2A in bladder cancer cells and overexpressed miR-381-3p could inhibit the progression of bladder cancer cells. MiR-381-3p/RAB2A axis activates cell proliferation and inhibits cell apoptosis in bladder cancer.

A Carotenoid- and Poly-ß-Hydroxybutyrate-Free Mutant Strain of Sphingomonas elodea ATCC 31461 for the Commercial Production of Gellan.

Gellan gum is a microbial exopolysaccharide, produced after aerobic fermentation using the Gram-negative bacterium strain Sphingomonas elodea ATCC 31461. Due to its unique structure and excellent physical characteristics, gellan gum has a broad range of applications in food, pharmaceutical, and other industries where it is used for stabilizing, emulsifying, thickening, and suspending. During the fermentative production of gellan, strain ATCC 31461 also accumulates large amounts of the metabolic by-products yellow carotenoid pigments and poly-ß-hydroxybutyrate (PHB), which is decreasing the gellan production and increasing processing costs. A pigment PHB-free mutant was obtained by knocking out the phytoene desaturase gene (crtI) in the carotenoid biosynthetic pathway and the phaC gene, encoding a PHB synthase for the polymerization of PHB. Unfortunately, the double gene knockout mutant produced only 0.56 g liter-1 gellan. Furthermore, blocking PHB and carotenoid synthesis resulted in the accumulation of pyruvate, which reduced gellan production. To elevate gellan production, combined UV irradiation and ethyl methanesulfonate (EMS) mutagenesis treatment were used. A mutant strain with the same level of pyruvate as that of the wild-type strain and higher gellan production was isolated (1.35 g liter-1, 132.8% higher than the double gene knockout mutant and 14.4% higher than the wild-type strain ATCC 31461). In addition, a new gellan gum recovery method based on the new mutant strain was investigated, in which only 30% isopropanol was required, which is twice for the wild-type strains, and the performance of the final product was improved. Thus, the mutant strain could be an ideal strain for the commercial production of gellan.IMPORTANCE A carotenoid- and PHB-free double gene knockout strain mutant was constructed to simplify the purification steps normally involved in gellan production. However, the production of gellan gum was unexpectedly reduced. A mutant with 14.4% higher gellan production than that of the wild-type strain was obtained and isolated after employing UV and EMS combined mutagenesis. Based on this high-yield and low-impurity-producing mutant, a new recovery method requiring less organic solvent and fewer operating steps was developed. This method will effectively reduce the production costs and improve the economic benefits of large-scale gellan production.

[The design of the software system for the interpretation and guidance of drug locus related to drug treatment].

Based on the pharmacogenomics theory, this study developed a software system for interpretation of drug gene loci and guidance on clinical safe medication with the purpose of providing clinical guidance on the safety and effectiveness of drug use through accurate and efficient detection and interpretation of drug gene loci. The system infrastructure was built on a service-oriented architecture (SOA) design and Docker container virtualization approach to achieve a rapid and automatic interpretation of genetic results and best available drugs. The front end was established on HTML5 and JavaScript to realize visualization of analysis results and user interaction. The system was tested and validated to show robust performance which is reliable in clinical use. It will show high impact on the development of pharmacogenomics and clinical practice of patients with personalized medicine.

Genome Sequence Resource for Ilyonectria mors-panacis, Causing Rusty Root Rot of Panax notoginseng.

Ilyonectria mors-panacis is the cause of a serious disease hampering the production of Panax notoginseng, an important Chinese medicinal herb, widely used for its anti-inflammatory, antifatigue, hepato-protective, and coronary heart disease prevention effects. Here, we report the first Illumina-Pacbio hybrid sequenced draft genome assembly of I. mors-panacis strain G3B and its annotation. The availability of this genome sequence not only represents an important tool toward understanding the genetics behind the infection mechanism of I. mors-panacis strain G3B but also will help illuminate the complexities of the taxonomy of this species.

Screening and enzymatic activity of high-efficiency gellan lyase producing bacteria Pseudoalteromonas hodoensis PE1.

Gellan is a widely used microbial polysaccharide and one of the more effective ways to expand its application value would be to investigate the mechanism of gellan lyase and to produce gellan oligosaccharide. In this study, efficient gellan degrading bacteria were screened. One of the strains with high efficient gellan degradation capacity was labeled PE1. Through physiological and biochemical analysis of 16S rDNA, the species was identified as Pseudoalteromonas hodoensis. The optimum conditions for enzymatic activity and how it was affected by metal ions were determined, and the results showed that the lyase activities were much higher than those of previously reported (about 20 times). The gellan degradation products were determined by thin-layer chromatography and the oligosaccharides were determined by high-efficiency liquid chromatography to analyze the action site of lyase. This study laid a solid foundation which elucidates the production and application of gellan oligosaccharides. Research highlights ● High efficiency gellan lyase producing bacteria ● Optimization of reaction conditions for gellan degradation ● Oligosaccharides were detected by TLC and HPLC to speculate the lyase action sites.

Design and synthesis of various quinizarin derivatives as potential anticancer agents in acute T lymphoblastic leukemia.

A series of quinizarin derivatives containing quaternary ammonium salts and/or thiourea groups were synthesized and their anticancer activities against leukemia cell lines have been tested. Results showed that most of quinizarin derivatives could inhibit the proliferation of leukemia cells. Among these derivatives, compound 3 showed good inhibition activity against various leukemia cells with IC50 values ranging from 0.90 ±â€¯2.55 µM to 10.90 ±â€¯3.66 µM. At the same time, compound 3 also inhibited the growth of human embryonic kidney-293 cell (HEK-293). Molt-4 and Jurkat cells, acute T lymphoblastic leukemia (T-ALL) cell lines, were selected to reveal potential anticancer mechanism of compound 3. Compound 3 inhibited the proliferation of Molt-4 and Jurkat cells in a dose- and time-dependent manner and led to a marked G0/G1 phase arrest. Analysis of Annexin V-APC and intracellular reactive oxygen species (ROS) level by flow cytometry showed that compound 3 induced significant apoptosis in Molt-4 and Jurkat cells. Western blotting assay showed that compound 3 activated the caspase-dependent apoptosis pathway and induced the degradation of Bcl-2 and c-myc protein.

Promotion of phenolic compounds production in Salvia miltiorrhiza hairy roots by six strains of rhizosphere bacteria.

Salvia miltiorrhiza Bunge is an important herb for the treatment of cerebrovascular and cardiovascular diseases with bioactive compounds (phenolic acids and tanshinones). Abundant studies showed that tanshinones could be stimulated by biotic and abiotic stresses, but limited information is available on biosynthesis of phenolic acids promoted by biotic stresses. The aim of the present work was to isolate and identify rhizosphere bacteria which stimulated phenolic compound in Salvia miltiorrhiza hairy roots and investigated the internal mechanism, providing a potential means to enhance content of pharmaceuticals in S. miltiorrhiza. The results showed that six bacteria, namely, HYR1, HYR26, SCR22, 14DSR23, DS6, and LNHR13, belonging to the genus Pseudomonas and Pantoea, significantly promoted the growth and content of major phenolic acids, RA and SAB. Bacteria LNHR13 was the most effective one, with the contents of RA and SAB reaching ∼2.5-fold (30.1 mg/g DW) and ∼2.3-fold (48.3 mg/g DW) as those of the control, respectively. Phytohormones and polysaccharides produced by bacteria showed potential responsibility for the growth and biosynthesis of secondary metabolites of S. miltiorrhiza. Meanwhile, we found that the more abundant the types and contents of phytohormones, the stronger their stimulating effect on the content of salvianolic acids.

Bacterial and diazotrophic diversities of endophytes in Dendrobium catenatum determined through barcoded pyrosequencing.

As an epiphyte orchid, Dendrobium catenatum relies on microorganisms for requisite nutrients. Metagenome pyrosequencing based on 16S rRNA and nifH genes was used to characterize the bacterial and diazotrophic communities associated with D. catenatum collected from 5 districts in China. Based on Meta-16S rRNA sequencing, 22 bacterial phyla and 699 genera were identified, distributed as 125 genera from 8 phyla and 319 genera from 10 phyla shared by all the planting bases and all the tissues, respectively. The predominant Proteobacteria varied from 71.81% (GZ) to 96.08% (YN), and Delftia (10.39-38.42%), Burkholderia (2.71-15.98%), Escherichia/Shigella (4.90-25.12%), Pseudomonas (2.68-30.72%) and Sphingomonas (1.83-2.05%) dominated in four planting bases. Pseudomonas (17.94-22.06%), Escherichia/Shigella (6.59-11.59%), Delftia (9.65-22.14%) and Burkholderia (3.12-11.05%) dominated in all the tissues. According to Meta-nifH sequencing, 4 phyla and 45 genera were identified, while 17 genera and 24 genera from 4 phyla were shared by all the planting bases and all the tissues, respectively. Burkholderia and Bradyrhizobium were the most popular in the planting bases, followed by Methylovirgula and Mesorhizobium. Mesorhizobium was the most popular in different tissues, followed by Beijerinckia, Xanthobacter, and Burkholderia. Among the genera, 39 were completely overlapped with the results based on the 16S rRNA gene. In conclusion, abundant bacteria and diazotrophs were identified in common in different tissues of D. catenatum from five planting bases, which might play a great role in the supply of nutrients such as nitrogen. The exact abundance of phylum and genus on the different tissues from different planting bases need deeper sequencing with more samples.

Synthesis, SAR and pharmacological characterization of novel anthraquinone cation compounds as potential anticancer agents.

Emodin, a natural anthraquinone derivative isolated from Rheum palmatum L., has been demonstrated to exhibit good anti-cancer effect. In this study, a series of novel quaternary ammonium salts of emodin, anthraquinone and anthrone were synthesized and their anticancer activities were tested in vitro. The effects of emodin quaternary ammonium salts on cell viability, apoptosis, intracellular ROS, and mitochondrial membrane potential were investigated in A375, BGC-823, HepG2 and HELF cells. The results demonstrated that compound 4a induced morphological changes and decreased cell viability. Apoptosis triggered by compound 4a was visualized using DAPI staining and Annexin V-FITC/PI staining. Compound 4a-induced apoptosis of A375 cells were showed to be associated with the dissipation of mitochondrial membrane potential (ΔΨm) as a result of the up-regulation of P53 and Caspase-3. When cancer cells were treated with emodin derivative, their ability to generate reactive oxygen species (ROS) rose significantly and the mitochondrial membrane potential decreased. Additionally, confocal microscopy assay confirmed that compound 4a was primarily located in the mitochondria of A375 cells. These results suggested that compound 4a has the potential for use in cancer therapy.

Unveiling of Dominant Fungal Pathogens Associated With Rusty Root Rot of Panax notoginseng Based on Multiple Methods.

Root rot is an important disease hampering the sustainable cultivation of Panax notoginseng. Culture-dependent and independent techniques were used to elucidate the dominant fungal pathogen of rusty root rot of P. notoginseng. Based on Illumina sequencing profiles for fungi using ITS primers, five phyla-namely Ascomycota, Basidiomycota, Glomeromycota, Zygomycota, and Chytridiomycota-were identified, and the analyses showed that the Ascomycota was the dominant phylum (∼50 to 97%), especially in the symptomatic samples. Out of 226 total genera identified, seven genera had over 1% average abundance, including Ilyonectria, Fusarium, Tetracladium, Cladosporium, Rhizophagus, Alternaria, and Perisporiopsis. However, only Ilyonectria was the predominant genera in the symptomatic samples (∼76 to 80%), while the others, including Fusarium, had higher abundances in asymptomatic samples. Based on in vitro and in vivo pathogenicity, the isolate G3B was demonstrated to be the pathogen causing rusty root rot of P. notoginseng, and it was identified as Ilyonectria mors-panacis. Based on primers F2-R2 targeting the His3 gene of Ilyonectria, real-time quantitative PCR (qPCR) was performed as an additional proof confirming that I. mors-panacis was the dominant pathogen in the symptomatic samples during the years of the study (2014-2015).