Engineering DMNT emission in cotton enhances direct and indirect defense against mirid bugs.

INTRODUCTION: As an important herbivore-induced plant volatile, (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) is known for its defensive role against multiple insect pests, including attracting natural enemies. A terpene synthase (GhTPS14) and two cytochrome P450 (GhCYP82L1, GhCYP82L2) enzymes are involved in the de novo synthesis of DMNT in cotton. We conducted a study to test the potential of manipulating DMNT-synthesizing enzymes to enhance plant resistance to insects. OBJECTIVES: To manipulate DMNT emissions in cotton and generate cotton lines with increased resistance to mirid bug Apolygus lucorum. METHODS: Biosynthesis and emission of DMNT by cotton plants were altered using CRISPR/Cas9 and overexpression approaches. Dynamic headspace sampling and GC-MS analysis were used to collect, identify and quantify volatiles. The attractiveness and suitability of cotton lines against mirid bug and its parasitoid Peristenus spretus were evaluated through various assays. RESULTS: No DMNT emission was detected in knockout CAS-L1L2 line, where both GhCYP82L1 and GhCYP82L2 were knocked out. In contrast, gene-overexpressed lines released higher amounts of DMNT when infested by A. lucorum. At the flowering stage, L114 (co-overexpressing GhCYP82L1 and GhTPS14) emitted 10-15-fold higher amounts than controls. DMNT emission in overexpressed transgenic lines could be triggered by methyl jasmonate (MeJA) treatment. Apolygus lucorum and its parasitoid were far less attracted to the double edited CAS-L1L2 plants, however, co-overexpressed line L114 significantly attracted bugs and female wasps. A high dose of DMNT, comparable to the emission of L114, significantly inhibited the growth of A. lucorum, and further resulted in higher mortalities. CONCLUSION: Turning down DMNT emission attenuated the behavioral preferences of A. lucorum to cotton. Genetically modified cotton plants with elevated DMNT emission not only recruited parasitoids to enhance indirect defense, but also formed an ecological trap to kill the bugs. Therefore, manipulation of DMNT biosynthesis and emission in plants presents a promising strategy for controlling mirid bugs.

Gut microbiome, T cell subsets, and cytokine analysis identify differential biomarkers in tuberculosis.

Introduction: The gut microbiota, T cell subsets, and cytokines participate in tuberculosis (TB) pathogenesis. To date, the mechanisms by which these factors interactively promote TB development at different time points remain largely unclear. In the context of this study, We looked into the microorganisms in the digestive tract, T cell types, and cytokines related to tuberculosis. Methods: According to QIIME2, we analyzed 16SrDNA sequencing of the gut microbiome on the Illumina MiSeq. Enzyme-linked immunosorbent assay was used to measure the concentrations of cytokines. Results: We showed the presence of 26 identifiable differential microbiomes in the gut and 44 metabolic pathways between healthy controls and the different time points in the development of TB in patients. Five bacterial genera (Bacteroides, Bifidobacterium, Faecalibacterium, Collinsella, and Clostridium) were most closely associated with CD4/CD8, whereas three bacterial taxa (Faecalibacterium, Collinsella, and Clostridium) were most closely associated with CD4. Three bacterial taxa (Faecalibacterium, Ruminococcus, and Dorea) were most closely associated with IL-4. Ruminococcus was most closely associated with IL-2 and IL-10. Conclusion: Diverse microorganisms, subsets of T cells, and cytokines, exhibiting varying relative abundances and structural compositions, were observed in both healthy controls and patients throughout distinct phases of tuberculosis. Gaining insight into the function of the gut microbiome, T cell subsets, and cytokines may help modulate therapeutic strategies for TB.

Prognostic Value of Hemoglobin Concentration on Renal Outcomes with Diabetic Kidney Disease: A Retrospective Cohort Study.

Objective: Diabetic kidney disease (DKD) patients with anemia face an elevated risk of glomerular filtration rate decline. However, the association between hemoglobin and estimated Glomerular Filtration Rate (eGFR) progression remains to be elucidated. Methods: A retrospective cohort of 815 subjects with DKD was followed from January 2010 to January 2023. A Cox proportional hazard regression model was utilized to explore the predictive role of hemoglobin in renal outcomes. Renal outcomes were defined as a composite endpoint, including a 50% decline in eGFR from baseline or progression to End-Stage Renal Disease (ESRD). To unveil any nonlinear relationship between hemoglobin and renal outcomes, Cox proportional hazard regression with cubic spline functions and smooth curve fitting was conducted. Additionally, subgroup analyses were performed to identify specific patient populations that might derive greater benefits from higher hemoglobin. Results: Among the 815 DKD subjects, the mean age was 56.482 ± 9.924 years old, and 533 (65.4%) were male. The mean hemoglobin was 121.521±22.960 g/L. The median follow-up time was 21.103±18.335 months. A total of 182 (22.33%) individuals reached the renal composite endpoint during the study period. After adjusting for covariates, hemoglobin was found to exert a negative impact on the renal composite endpoint in patients with DKD (HR 0.975, 95% CI [0.966, 0.984]). A nonlinear relationship between hemoglobin and the renal composite endpoint was identified with an inflection point at 109 g/L. Subgroup analysis unveiled a more pronounced association between hemoglobin and renal prognosis in males. Conclusion: Hemoglobin emerges as a predictive indicator for the renal prognosis of diabetic kidney disease in China. This study reveals a negative and non-linear relationship between hemoglobin levels and the renal composite endpoint. A substantial association is noted when hemoglobin surpasses 109 g/L in relation to the renal composite endpoint.

Rapid seedling emergence of invasive Phytolacca americana is related to higher soluble sugars produced by starch metabolism and photosynthesis compared to native P. acinosa.

Seedling emergence is an essential event in the life cycle of plants. Most invasive plants have an advantage in population colonization over native congeners. However, differential seedling emergence between invasive plants and native congeners, especially their mechanisms, have rarely been explored. In this study, we show that the seedlings of invasive Phytolacca americana emerge faster compared to native P. acinosa. Genome-wide transcriptomes of initially germinated seeds versus seedlings at 4 days after germination (DAG) suggested that differentially expressed genes (DEGs) in the photosynthesis-antenna proteins pathway were up-regulated in both P. americana and P. acinosa, while DEGs in starch and sucrose metabolism were significantly down-regulated in P. americana. Gene expression analysis indicated that photosynthesis-related DEGs reached their highest level at 3 DAG in P. americana, while they peaked at 4 DAG in P. acinosa. We also identified one ß-amylase gene in P. americana (PameAMYB) that showed the highest expression at 1 DAG, and two ß-amylase genes in P. acinosa that expressed lower than PameAMYB at 0 and 1 DAG. Enzymatic activity of ß-amylases also suggested that P. americana had the highest activity at 1 DAG, which was earlier than P. acinosa (at 4 DAG). Soluble sugars, the main source of energy for seedling emergence, were showed higher in P. americana than in P. acinosa, and reached the highest at 4 DAG that positively affected by photosynthesis. These results indicate that the rapid seedling emergence of invasive P. americana benefited from the high soluble sugar content produced by starch metabolism and photosynthesis. Altogether, this work contributes to our fundamental knowledge on physiological and molecular mechanisms for plant invasion success.

Revitalizing antitumor immunity: Leveraging nucleic acid sensors as therapeutic targets.

Nucleic acid sensors play a critical role in recognizing and responding to pathogenic nucleic acids as danger signals. Upon activation, these sensors initiate downstream signaling cascades that lead to the production and release of pro-inflammatory cytokines, chemokines, and type I interferons. These immune mediators orchestrate diverse effector responses, including the activation of immune cells and the modulation of the tumor microenvironment. However, careful consideration must be given to balancing the activation of nucleic acid sensors to avoid unwanted autoimmune or inflammatory responses. In this review, we provide an overview of nucleic acid sensors and their role in combating cancer through the perception of various aberrant nucleic acids and activation of the immune system. We discuss the connections between different programmed cell death modes and nucleic acid sensors. Finally, we outline the development of nucleic acid sensor agonists, highlighting how their potential as therapeutic targets opens up new avenues for cancer immunotherapy.

Predictive performance of triglyceride glucose index (TyG index) to identify glucose status conversion: a 5-year longitudinal cohort study in Chinese pre-diabetes people.

OBJECTIVE: Triglyceride glucose index (TyG index) has been recommended as an alternative indicator of insulin resistance. However, the association between TyG and regression from prediabetes to normoglycemia remains to be elucidated. METHODS: This retrospective cohort study involved 25,248 subjects with prediabetes at baseline conducted from 2010 to 2016. A Cox proportional hazard regression model was designed to evaluate the role of TyG in identifying people at converting from prediabetes to normoglycemia. Cox proportional hazards regression with cubic spline functions and smooth curve fitting was used to dig out the nonlinear relationship between them. Detailed evaluations for TyG were also performed using sensitivity and subgroup analyse. RESULTS: Among the included prediabetes subjects (n = 25,248), the mean age was 49.27 ± 13.84 years old, and 16,701 (66.15%) were male. The mean TyG was 8.83 ± 0.60. The median follow-up time was 2.96 ± 0.90 years. 11,499 (45.54%) individuals had a final diagnosis of normoglycemia. After adjusting for covariates, TyG was negatively affecting the results of glucose status conversion in prediabetes people (HR 0.895, 95% CI 0.863, 0.928). There was a nonlinear connection between TyG and normoglycemia in prediabetes people, and the inflection point was 8.88. The effect sizes (HR) on the left and right sides of the inflection point were 0.99 (0.93, 1.05) and 0.79 (0.74, 0.85), respectively. Sensitivity analysis confirmed the robustness of these results. Subgroup analysis showed that TyG was more strongly associated with incident glucose status conversion in male, BMI ≥ 25. In contrast, there was a weaker relationship in those with female, BMI < 25. CONCLUSION: Based on sample of subjects evaluated between 2010 and 2016, TyG index appears to be a promising marker for predicting normoglycemic conversion among prediabetes people in China. This study demonstrates a negative and non-linear association between TyG and glucose status conversion from prediabetes to normoglycemia. TyG is strongly related to glucose status conversion when TyG is above 8.88. From a therapeutic point of view, it is meaningful to maintain TyG levels within the inflection point to 8.88.

Enhanced single-base mutation diversity by the combination of cytidine deaminase with DNA-repairing enzymes in yeast.

The occurrence of random mutations can increase the diversity of the genome and promote the evolutionary process of organisms. High efficiency mutagenesis techniques significantly accelerate the evolutionary process. In this work, we describe a targeted mutagenesis system named MutaT7trans to significantly increase mutation rate and generate mutations across all four nucleotides in yeast. We constructed different DNA-repairing enzyme-PmCDA1-T7 RNA polymerase (T7 RNAP) fusion proteins, achieved targeted mutagenesis by flanking the target gene with T7 promoters, and tuned the mutation spectra by introducing different DNA-repairing enzymes. With this mutagenesis tool, the proportion of non-C â†’ T mutations was 10-11-fold higher than the cytidine deaminase-based evolutionary tools, and the transversion mutation frequency was also elevated. The mutation rate of the target gene was significantly increased to 5.25 × 10-3 substitutions per base (s. p. b.). We also demonstrated that MutaT7trans could be used to evolve the CrtE, CrtI, and CrtYB gene in the ß-carotene biosynthesis process and generate different types of mutations.

Ejection dynamics of spherically confined active polymers through a small pore.

Using Brownian dynamics simulations, we study the ejection dynamics of spherically confined active polymers through a small pore. Although the active force can provide a driving force other than the entropy drive, it also causes the collapse of the active polymer, which in turn reduces the entropy drive. Thus, our simulation results confirm that the active polymer's ejection process can be divided into three stages. In the first stage, the influence of the active force is small, and the ejection is mainly an entropy-driven process. In the second stage, the ejection time satisfies the scaling relationship with the chain length, and the value of obtained scaling exponent is less than 1.0, indicating that the active force accelerates the ejection process. In the third stage, the scaling exponent is maintained at about 1.0, where the active force dominates the ejection process, and the ejection time is inversely proportional to the Péclet number. Furthermore, we find that the ejection velocity of the trailing particles has significant differences at different stages and is the core factor of the ejection mechanism at different stages. Our work helps us understand this non-equilibrium dynamic process and enhances our prediction of the relevant physiological phenomena.

Conformation of a Comb-like Chain in Solution: Effect of Backbone Rigidity.

We study the effect of backbone rigidity on the conformation of comb-like chains in dilute solution by using Brownian dynamics simulations. Our results demonstrate that the backbone rigidity can control the effect of side chains on the conformation of comb-like chains; that is, the relative strength of the excluded-volume interactions from backbone monomer-graft and graft-graft to backbone monomer-monomer gradually weakens with the increase of backbone rigidity. Only when the rigidity of the backbone tends to be flexible and the grafting density is high is the effect of excluded volume of graft-graft on the conformation of comb-like chains significant enough, and other cases can be ignored. Our results show that the radius of gyration of comb-like chains and the persistence length of the backbone are exponentially related to the stretching factor, where the power exponent exhibits an increase with the increase of the strength of bending energy. These finds provide new insights for characterizing the structure properties of comb-like chains.

Heat Shock Protein Inhibitors Show Synergistic Antibacterial Effects with Photodynamic Therapy on Caries-Related Streptococci In Vitro and In Vivo.

Caries are chronic infections in which the cariogenic biofilm plays a critical role in disease occurrence and progression. Photodynamic therapy (PDT) is a new effective treatment that is receiving wide attention in the antibacterial field, but it can lead to the upregulation of heat shock proteins (HSPs), which enhances bacterial resistance. Herein, we incorporated HSP inhibitors with PDT to evaluate the effect on Streptococcus mutans, Streptococcus sobrinus, and Streptococcus sanguinis under planktonic conditions and on cariogenic biofilms. Additionally, a model of caries was established in 2-week-old rats, and anticaries properties were evaluated by Keyes' scoring. Importantly, the combination of HSP inhibitors and PDT had outstanding efficiency in inhibiting the growth of tested Streptococcus strains and the formation of either monomicrobial or multispecies biofilms in vitro. In addition, the quantity of colonized streptococci and the severity of carious lesions were also distinctly suppressed in vivo. Overall, the synergistic application of HSP inhibitors and PDT has promising potential in the prevention and treatment of dental caries. IMPORTANCE Effective therapies for the prevention and control of caries are urgently needed. Cariogenic streptococci play a key role in the occurrence and progression of caries. Recently, photodynamic therapy has been demonstrated to have good antibacterial efficiency, but it can cause a heat shock response in bacteria, which may weaken its practical effects. We indicate here an effective therapeutic strategy of combining heat shock protein inhibitors and photodynamic therapy, which shows excellent inhibition toward three dominant streptococci related to caries and suppression of carious progression in a rat model. Further development for clinical application is promising.

Profiling the oral microbiomes in patients with Alzheimer's disease.

OBJECTIVES: To analyse the characteristics of the oral microbiomes and expected to find biomarkers about Alzheimer's disease (AD). SUBJECTS AND METHODS: AD patients (n = 26) and cognitive intact people (n = 26) were examined for cognition, depression, oral health and collected saliva and gingival crevicular fluid (GCF) in the morning. Full-length 16S rRNA gene was amplified and sequencing was performed using the PacBio platform. RESULTS: The predominant bacterium of salivary microbiome and periodontal microbiome from AD patients was Streptococcus oralis and Porphyromonas gingivalis, respectively. With respect to ß diversity analysis, there was a significance difference in periodontal microbiome between AD patients and cognitively intact subjects. The relative abundance of Veillonella parvula significantly increased in oral microbiomes from AD patients. Interestingly, the dominant species were different between early-onset AD and late-onset AD patients. Moreover, the predominant species were changed as the clinical severity of AD. Furthermore, the correlation analysis revealed that V. parvula was associated with AD in both saliva and GCF and that P. gingivalis was associated with AD only in GCF. CONCLUSIONS: In this study, the microbiome community of oral microbes was altered in AD patients and periodontal microbiome was sensitive to cognition changes. Moreover, V. parvula and P. gingivalis were associated with AD.

Invasive Plants Have Higher Resistance to Native Generalist Herbivores Than Exotic Noninvasive Congeners.

Research on the invasive plant Phytolacca americana (L.) mostly focuses on its medicinal value and enrichment of heavy metals. However, little is known regarding its impact on native herbivorous insects. In this study, we explored the effects of P. americana and the exotic noninvasive Phytolacca icosandra (L.) on the Spodoptera litura (Fabricius) (native tobacco cutworm) via bioassay, oviposition preference, detoxifying enzyme activity analysis, and phytochemical determination. We found that the oviposition preference index (OPI) of S. litura feeding on P. icosandra was higher than that of P. americana. The developmental duration of S. litura feeding on P. icosandra was shorter than that of P. americana. Additionally, the Acetylcholinesterase (AchE) and Glutathione-S-transferase (GST) activities of S. litura feeding on P. americana were higher than that of S. litura feeding on artificial diets or P. icosandra. The content of lignin and flavonoids in P. americana was relatively high, whereas starch content was relatively low. These findings suggest invasive plants have higher resistance to herbivores, thereby suffering less damage than exotic noninvasive plants.

Effect of plant VOCs and light intensity on growth and reproduction performance of an invasive and a native Phytolacca species in China.

Invasive plants often pose great threats to the growth of co-occurring native plant species. Identifying environmental factors that facilitate exotic plant invasion and native species decline are important. In this study, we measured the effects of plant volatile organic compounds (VOCs), light intensity, and their interactions on the growth and reproduction performance of indigenous Phytolacca acinosa, and invasive Phytolacca americana, which has largely replaced the former in China. VOCs of invasive P. americana and low light levels both had negative effects on P. acinosa morphological and reproductive traits (stem length, average leaf number, total number, and length of racemes), and biomass allocation (total biomass, and leaf and flower mass fraction); low light also affected photosynthesis-related trait (specific leaf area) of P. acinosa. In contrast, VOCs of P. acinosa had no significant effect on P. americana, but low light levels adversely affected its morphological and reproductive traits (stem length, total number, and length of racemes) and biomass allocation (total biomass, stem, and leaf mass fraction). Interactions between plant VOCs and light intensity had no significant effects on P. acinosa or P. americana. Under all experimental treatments, stem length, average leaf area, total number, and length of racemes, Root/Shoot ratio, root and flower mass fraction of P. americana were higher than those of P. acinosa, while average leaf number, specific leaf area, and leaf mass fraction was lower. These results indicated that P. acinosa was sensitive to P. americana VOCs and low light, which might affect the growth of sympatric P. acinosa. P. americana was negatively influenced by low light, but higher plant height and more reproductive organ resource allocation relative to sympatric P. acinosa might contribute to invasion success.

Bioresponsive nanotherapy for preventing dental caries by inhibiting multispecies cariogenic biofilms.

Early childhood caries (ECC) is a public healthcare concern that greatly reduces the quality of life of young children. As a leading factor of ECC, cariogenic biofilms are composed of acidogenic/aciduric pathogens and extracellular polysaccharides (EPSs), creating an acidic and protected microenvironment. Antimicrobial photodynamic therapy (aPDT) is a noninvasive, painless, and efficient therapeutic approach that is suitable for treating ECC. However, due to the hyperfine structure of cariogenic biofilms, most photosensitizers (PSs) could not access and penetrate deeply in biofilms, which dramatically hamper their efficiency in the clinic. Herein, bioresponsive nanoparticle loaded with chlorin e6 (MPP-Ce6) is developed, which largely increases the penetration depth (by over 75%) and retention (by over 100%) of PS in the biofilm compared with free Ce6. Furthermore, MPP-Ce6-mediated aPDT not only kills the bacteria in preformed biofilms but also inhibits multispecies biofilm formation. A rampant caries model is established to mimic ECC in vivo, where the population of cariogenic bacteria is decreased to 10% after MPP-Ce6-mediated aPDT. Importantly, the number and severity of carious lesions are efficiently reduced via Keyes' scoring and micro-CT analysis. This simple but effective strategy can serve as a promising approach for daily oral hygiene in preventing ECC.

Horizontal Spatial Metaphor Representation of Social Status in Chinese Culture.

The mapping relationship between social status and horizontal space (left/right) in Chinese culture has a long history. In order to explore the representation pattern of horizontal spatial metaphor of social status in Chinese culture, this study introduced two direct measurements, implicit relational assessment procedure (IRAP) paradigm, and spatial placement task to evaluate the mapping of social status to horizontal space. A total of 144 Chinese undergraduates participated in the research, wherein they were asked to place certain words indicating social status in either left or right box before or after the IRAP computer test. The results from the two measurements consistently showed that the mode of HIGH SOCIAL STATUS-LEFT and LOW SOCIAL STATUS-RIGHT (HLLR) had an advantage over HIGH SOCIAL STATUS-RIGHT and LOW SOCIAL STATUS-LEFT (HRLL), implying that the representation pattern of horizontal spatial metaphor of social status for the Chinese is HIGH SOCIAL STATUS-LEFT and LOW SOCIAL STATUS-RIGHT. However, the result convergence of the two measurements was not high, which suggests that embodiment effect has multiple characteristics and new specific experimental paradigms should be created to measure it.

Hoyosella suaedae sp. nov., a novel bacterium isolated from rhizosphere soil of Suaeda aralocaspica (Bunge) Freitag & Schütze.

A Gram-stain-positive, non-motile and coccus-shaped bacterium, designated strain LNNU 331112T, was isolated from the composite rhizosphere soil of the halophyte Suaeda aralocaspica (Bunge) Freitag and Schütze, which was collected in Xinjiang, north-west China. Growth occurred at 10-45 °C, pH 6.0-11.0 and in the presence of 0-10 % NaCl (w/v). Phylogenetic analysis based on the 16S rRNA gene sequence suggested that strain LNNU 331112T belonged to the genus Hoyosella and showed 95.6, 95.5 and 95.4 % sequence similarities to Hoyosella altamirensis DSM 45258T, Hoyosella subflava CGMCC 4.3532T and Hoyosella rhizosphaerae CGMCC 1.15478T, respectively. The estimated digital DNA-DNA hybridization relatedness values between strain LNNU 331112T and the type strains of H. altamirensis DSM 45258T, H. subflava CGMCC 4.3532T and H. rhizosphaerae CGMCC 1.15478T were 18.9, 19.3 and 18.3 %, respectively. The average nucleotide identity values between strain LNNU 331112T and H. altamirensis DSM 45258T, H. subflava CGMCC 4.3532T and H. rhizosphaerae CGMCC 1.15478T were 72.6, 72.7 and 72.3 %, respectively. The genome sequence of strain LNNU 331112T showed 69.0-72.3 % average amino acid identity values in comparison with the related genome sequences of three validly published Hoyosella species. The genome of strain LNNU 331112T was 3.47 Mb, with a DNA G+C content of 68.4 mol%. A total of 3182 genes were identified as protein-coding in strain LNNU 331112T. Genomic analysis revealed that a number of genes involved in osmotic pressure regulation, intracellular pH homeostasis and potassium (K+) uptake protein were found in strain LNNU 331112T. The predominant menaquinones were MK-8 (44.6 %) and MK-7 (55.4 %), which differentiated strain LNNU 331112T from other three recognized Hoyosella species. Major fatty acids (>10 %) were C17 : 1 ω8c (33.8 %), C16 : 0 (23.3 %), C17 : 0 (12.8 %) and summed feature 3 (12.9 %), which also clearly separated strain LNNU 331112T from three recognized Hoyosella species. The polar lipid profile of strain LNNU 331112T included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, one unidentified glycolipid, one unidentified phospholipid and two unidentified lipids. According to the results of phenotypic, chemotaxonomic and phylogenetic analyses, strain LNNU 331112T is considered to represent a novel species of the genus Hoyosella, for which the name Hoyosella suaedae sp. nov. is proposed. The type strain is LNNU 331112T (=KCTC 39808T=CGMCC 1.17107T=DSM 103463T).

Ruania rhizosphaerae sp. nov., a novel actinobacterium isolated from rhizosphere of Suaeda aralocaspica.

A Gram-stain-positive, non-motile and short rod-shaped actinobacterium, designated strain LNNU 22110T, was isolated from the rhizosphere soil of the halophyte Suaeda aralocaspica (Bunge) Freitag and Schütze, which collected in Xinjiang, north-west China. Growth occurred at 10-45 °C, pH 6.0-10.0 and in the presence of 0-11 % NaCl (w/v). Based on the results of 16S rRNA gene sequence phylogenetic analyses, strain LNNU 22110T belonged to the genus Ruania and had 97.5 and 95.5 % sequence similarity to Ruania alba KCTC 19413T and Ruania albidiflava CGMCC 4.3142T, respectively. The digital DNA-DNA hybridization relatedness values between strain LNNU 22110T and R. alba KCTC 19413T and R. albidiflava CGMCC 4.3142T were 23.2 and 19.9 %, respectively. The highest average nucleotide identity value between strain LNNU 22110T and its closest related strain (R. alba KCTC 19413T) was 80.2 %, much lower than the species delineation threshold of 95-96 %. The genome of strain LNNU 22110T was 4.4 Mb, with a genomic DNA G+C content of 68.4 mol%. The diagnostic diamino acids in the peptidoglycan layer of strain LNNU 22110T were lysine, alanine, glycine, glutamic acid and aspartic acid. The predominant menaquinone was MK-8(H4). The major fatty acid (>10 %) was anteiso-C15 : 0. The polar lipid profile of strain LNNU 22110T included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, diacylated phosphatidyl dimannoside, one unidentified glycolipid and two unidentified phospholipids. According to the phenotypic, phylogenetic and chemotaxonomic results, strain LNNU 22110T is considered to represent a novel species of the genus Ruania, for which the name Ruania rhizosphaerae sp. nov. is proposed. The type strain is LNNU 22110T (=KCTC 39807T=CGMCC 1.17105T).

Engineering Nicotiana tabacum for the de novo biosynthesis of DMNT to regulate orientation behavior of the parasitoid wasps Microplitis mediator.

BACKGROUND: (E)-4,8-dimethylnona-1,3,7-triene (DMNT), one of the homoterpenes, is thought to contribute to plant indirect defense against insect herbivores. DMNT-enriched plants have great application potential to regulate insect behavior in the 'push & pull' strategy of pest management. However, de novo biosynthesis of DMNT in plants without a homoterpene metabolic pathway in their wild type is still not achieved, and the role of DMNT played in these plants and their interacted insects remains unclear. RESULTS: Cytochrome P450s and terpene synthases involved in homoterpenes biosynthesis in cotton plants were employed to generate DMNT-releasing tobacco plants. Single GhTPS14 transgenic Nicotiana tabacum only emitted (E)-nerolidol, the precursor of DMNT. Transgenic tobaccos expressing single GhCYP82Ls were unable to produce DMNT or TMTT, while DMNT was detected when exogenous (E)-nerolidol was added. Compared to wild-type plants, only co-expression of GhCYP82Ls and GhTPS14 in transgenic tobaccos triggered the constitutive release of single-component DMNT. Furthermore, DMNT-emitting transgenic tobacco plants, whether infested with Helicoverpa armigera larvae or not, significantly incited orientation behavior of parasitoid wasps Microplitis mediator. CONCLUSION: Wild type N. tabacum plants have no DMNT metabolic pathway. DMNT could be de novo biosynthesized via co-expression of GhCYP82Ls and GhTPS14. What is more, the parasitoid wasp M. mediator could be recruited by DMNT-releasing transgenic tobaccos, especially by H. armigera-infested transgenic tobaccos, suggesting the potential roles of engineered N. tabacum in regulating the behavioral preference of M. mediator.

Full-length transcriptome analysis of Phytolacca americana and its congener P. icosandra and gene expression normalization in three Phytolaccaceae species.

BACKGROUND: Phytolaccaceae species in China are not only ornamental plants but also perennial herbs that are closely related to human health. However, both large-scale full-length cDNA sequencing and reference gene validation of Phytolaccaceae members are still lacking. Therefore, single-molecule real-time sequencing technology was employed to generate full-length transcriptome in invasive Phytolacca americana and non-invasive exotic P. icosandra. Based on the transcriptome data, RT-qPCR was employed to evaluate the gene expression stability in the two plant species and another indigenous congener P. acinosa. RESULTS: Total of 19.96 Gb and 19.75 Gb clean reads of P. americana and P. icosandra were generated, including 200,857 and 208,865 full length non-chimeric (FLNC) reads, respectively. Transcript clustering analysis of FLNC reads identified 89,082 and 98,448 consensus isoforms, including 86,989 and 96,764 high-quality ones. After removing redundant reads, 46,369 and 50,220 transcripts were obtained. Based on structure analysis, total 1675 and 1908 alternative splicing variants, 25,641 and 31,800 simple sequence repeats (SSR) as well as 34,971 and 36,841 complete coding sequences were detected separately. Furthermore, 3574 and 3833 lncRNA were predicted and 41,676 and 45,050 transcripts were annotated respectively. Subsequently, seven reference genes in the two plant species and a native species P. acinosa were selected and evaluated by RT-qPCR for gene expression analysis. When tested in different tissues (leaves, stems, roots and flowers), 18S rRNA showed the highest stability in P. americana, whether infested by Spodoptera litura or not. EF2 had the most stable expression in P. icosandra, while EF1-α was the most appropriate one when attacked by S. litura. EF1-α showed the highest stability in P.acinosa, whereas GAPDH was recommended when infested by S. litura. Moreover, EF1-α was the most stable one among the three plant species whenever germinating seeds or flowers only were considered. CONCLUSION: Full-length transcriptome of P. americana and P. icosandra were produced individually. Based on the transcriptome data, the expression stability of seven candidate reference genes under different experimental conditions was evaluated. These results would facilitate further exploration of functional and comparative genomic studies in Phytolaccaceae and provide insights into invasion success of P. americana.

Effect of Ag-1031 on apoptosis in gastric cancer AGS cells and its effects on the PI3K/AKT/mTOR signaling pathway.

PURPOSE: To investigate the effect of Ag-1031 on apoptosis of gastric cancer AGS cells and the PI3K/AKT/mTOR signaling pathway. METHODS: Gastric cancer cells were treated with different doses of Ag-1031. Cell viability was determined with MTT assay. Apoptosis was analyzed with Hoechst 33342 staining and Annexin V-FITC/PI double staining. The protein expression levels of p-PI3K, p-AKT, p-mTOR, PI3K, AKT and mTOR were assayed with western blotting method. RESULTS: Different doses of Ag-1031 significantly inhibited the proliferation of AGS cells. Moreover, Ag-1031 induced apoptosis,in gastric cancer cells. In addition, Ag-1031 significantly and dose-dependently inhibited the protein expressions of p-PI3K, p-AKT and p-mTOR. However, it had no effect on the protein expressions of PI3K, AKT and mTOR. CONCLUSION: In this study, it was found for the first time that Ag-1031 exerts an anti-tumor effect by induction apoptosis in gastric cancer AGS cells, through a mechanism related to inhibition of the activation of the PI3K/AKT/mTOR signaling pathway.