Flexible Luminescent Hydrogen-bonded Organic Framework for the Separation of Benzene and Cyclohexane.

A nonplanar phenothiazine derivative with three cyano moieties (PTTCN) is designed and synthesized to achieve functional crystals for absorptive separation of benzene and cyclohexane. PTTCN can crystallize into two kinds of crystals with different fluorescence colors in different solvent systems. The molecules in two crystals are in different stereo isomeric forms of nitrogen, quasi axial (ax), and quasi equatorial (eq). The crystals with blue fluorescence in ax form may selectively adsorb benzene by a single-crystal-to-single-crystal (SCSC) transformation, but separated benzene from a benzene/cyclohexane equimolar mixture with a low purity of 79.6%. Interestingly, PTTCN molecules with eq form and benzene co-assembled to construct a hydrogen-bonded framework (X-HOF-4) with S-type solvent channels and yellow-green fluorescence, and can release benzene to form nonporous guest-free crystal under heating. Such nonporous crystals strongly favor aromatic benzene over cyclohexane and may selectively reabsorb benzene from benzene/cyclohexane equimolar mixture to recover original framework, and the purity of benzene can reach ≈96.5% after release from framework. Moreover, reversible transformation between the nonporous crystals and the guest-containing crystals allows the material to be reused.

Niche-Specific Restructuring of Bacterial Communities Associated with Submerged Macrophyte under Ammonium Stress.

Submerged macrophytes and their epiphytic microbes form a "holobiont" that plays crucial roles in regulating the biogeochemical cycles of aquatic ecosystems but is sensitive to environmental disturbances such as ammonium loadings. Increasingly more studies suggest that plants may actively seek help from surrounding microbial communities whereby conferring benefits in responding to particular abiotic stresses. However, empirical evidence is scarce regarding how aquatic plants reconstruct their microbiomes as a "cry-for-help" against acute ammonium stress. Here, we investigated the temporal dynamics of the phyllosphere and rhizosphere bacterial communities of Vallisneria natans following ammonium stress and recovery periods. The bacterial community diversity of different plant niches exhibited opposite patterns with ammonium stress, that is, decreasing in the phyllosphere while increasing in the rhizosphere. Furthermore, both phyllosphere and rhizosphere bacterial communities underwent large compositional changes at the end of ammonium stress, significantly enriching of several nitrifiers and denitrifiers. Meanwhile, bacterial legacies wrought by ammonium stress were detected for weeks; some plant growth-promoting and stress-relieving bacteria remained enriched even after stress disappeared. Structural equation model analysis showed that the reshaped bacterial communities in plant niches collectively had a positive effect on maintaining plant biomass. Additionally, we applied an age-prediction model to predict the bacterial community's successional trajectory, and the results revealed a persistent change in bacterial community development under ammonium treatment. Our findings highlight the importance of plant-microbe interactions in mitigating plant stress and fostering a better understanding of the assembly of plant-beneficial microbes under ammonium stress in aquatic ecosystems. IMPORTANCE Increasing anthropogenic input of ammonium is accelerating the decline of submerged macrophytes in aquatic ecosystems. Finding efficient ways to release submerged macrophytes from ammonium stress is crucial to maintain their ecological benefits. Microbial symbioses can alleviate abiotic stress in plants, but harnessing these beneficial interactions requires a detailed understanding of plant microbiome responses to ammonium stress, especially over a continuous time course. Here, we tracked the temporal changes in bacterial communities associated with the phyllosphere and rhizosphere of Vallisneria natans during ammonium stress and recovery periods. Our results showed that severe ammonium stress triggers a plant-driven timely reshaping of the associated bacterial community in a niche-specific strategy. The reassembled bacterial communities could potentially benefit the plant by positively contributing to nitrogen transformation and plant growth promotion. These findings provide empirical evidence regarding the adaptive strategy of aquatic plants whereby they recruit beneficial microbes against ammonium stress.

Lower Compositional Variation and Higher Network Complexity of Rhizosphere Bacterial Community in Constructed Wetland Compared to Natural Wetland.

Macrophyte rhizosphere microbes, as crucial components of the wetland ecosystem, play an important role in maintaining the function and stability of natural and constructed wetlands. Distinct environmental conditions and management practices between natural and constructed wetlands would affect macrophytes rhizosphere microbial communities and their associated functions. Nevertheless, the understanding of the diversity, composition, and co-occurrence patterns of the rhizosphere bacterial communities in natural and constructed wetlands remains unclear. Here, we used 16S rRNA gene high-throughput sequencing to characterize the bacterial community of the rhizosphere and bulk sediments of macrophyte Phragmites australis in representative natural and constructed wetlands. We observed higher alpha diversity of the bacterial community in the constructed wetland than that of the natural wetland. Additionally, the similarity of bacterial community composition between rhizosphere and bulk sediments in the constructed wetland was increased compared to that of the natural wetland. We also found that plants recruit specific taxa with adaptive functions in the rhizosphere of different wetland types. Rhizosphere samples of the natural wetland significantly enriched the functional bacterial groups that mainly related to nutrient cycling and plant-growth-promoting, while those of the constructed wetland-enriched bacterial taxa with potentials for biodegradation. Co-occurrence network analysis showed that the interactions among rhizosphere bacterial taxa in the constructed wetland were more complex than those of the natural wetland. This study broadens our understanding of the distinct selection processes of the macrophytes rhizosphere-associated microbes and the co-occurrence network patterns in different wetland types. Furthermore, our findings emphasize the importance of plant-microbe interactions in wetlands and further suggest P. australis rhizosphere enriched diverse functional bacteria that might enhance the wetland performance through biodegradation, nutrient cycling, and supporting plant growth.

Mutation of OsLPR3 Enhances Tolerance to Phosphate Starvation in Rice.

Low Phosphate Root (LPR) encodes a protein localized to the endoplasmic reticulum (ER) and cell wall. This gene plays a key role in responding to phosphate (Pi) deprivation, especially in remodeling the root system architecture (RSA). An identification and expression analysis of the OsLPR family in rice (Oryza sativa) has been previously reported, and OsLPR5, functioning in Pi uptake and translocation, is required for the normal growth and development of rice. However, the role of OsLPR3, one of the five members of this family in rice, in response to Pi deficiency and/or in the regulation of plant growth and development is unknown. Therefore, in this study, the roles of OsLPR3 in these processes were investigated, and some functions were found to differ between OsLPR3 and OsLPR5. OsLPR3 was found to be induced in the leaf blades, leaf sheaths, and roots under Pi deprivation. OsLPR3 overexpression strongly inhibited the growth and development of the rice but did not affect the Pi homeostasis of the plant. However, oslpr3 mutants improved RSA and Pi utilization, and they exhibited a higher tolerance to low Pi stress in rice. The agronomic traits of the oslpr3 mutants, such as 1000-grain weight and seed length, were stimulated under Pi-sufficient conditions, indicating that OsLPR3 plays roles different from those of OsLPR5 during plant growth and development, as well as in the maintenance of the Pi status of rice.

Distribution of phenanthrene in the ospho2 reveals the involvement of phosphate on phenanthrene translocation and accumulation in rice.

The intricate mechanisms involved in the acquisition and translocation of polycyclic aromatic hydrocarbons (PAHs) in plants have not been elucidated. Phosphate (Pi) is the bioavailable form of essential macronutrient phosphorus, which is acquired and subsequently assimilated for plant optimal growth and development. Rice phosphate overaccumulator 2 (OsPHO2) is a central constituent of the regulation of Pi homeostasis in rice. In the present study, the role of OsPHO2 in regulating the translocation and accumulation of phenanthrene (Phe) and the involvement of Pi in this process were investigated. The temporal study (1 d-35 d) revealed a significant and gradual increase of Phe accumulation in Pi-deprived roots of wild-type (WT) seedlings. Compared with the WT, the concentrations of Phe were significantly higher in the shoots of ospho2 (OsPHO2 mutant) grown hydroponically with Phe (1.5 mg/L) under +Pi (200 µM) and -Pi (10 µM) conditions. The sap experiment clearly showed the significant increases in levels of Phe in the xylem sap of ospho2 than the WT grown hydroponically with Phe and +Pi. Further, the concentrations of both Phe and P were coordinately higher in the culms and flag leaves of the mutants than WT at maturity in potting soil with LPhe (6 mg/kg) and HPhe (60 mg/kg). However, the concentrations of Phe in the seeds were comparable in the WT and mutants, suggesting a pivotal of OsPHO2 in attenuating Phe toxicity in the seed. In +Phe WT, the relative expression level of OsPHO2 in the shoots was significantly lower, while those of Pi transporters (PTs) OsPT4 and OsPT8 were significantly higher in the roots compared with -Phe. Together, the results provided evidence towards the involvement of Pi in OsPHO2-regulated translocation and accumulation of Phe in rice.

OsSQD1 at the crossroads of phosphate and sulfur metabolism affects plant morphology and lipid composition in response to phosphate deprivation.

In phosphate (Pi)-deprived Arabidopsis (Arabidopsis thaliana), phosphatidylglycerol (PG) is substituted by sulfolipid for maintaining Pi homeostasis. Sulfoquinovosyl diacylglycerol1 (AtSQD1) encodes a protein, which catalyzes uridine diphosphate glucose (UDPG) and sulfite (SO32- ) to UDP-sulfoquinovose, which is a key component in the sulfolipid biosynthetic pathway. In this study, a reverse genetics approach was employed to decipher the function of the AtSQD1 homolog OsSQD1 in rice. Differential expressions of OsSQD1 in different tissue and response to -P and -S also detected, respectively. The in vitro protein assay and analysis suggests that OsSQD1 is a UDP-sulfoquinovose synthase. Transient expression analysis showed that OsSQD1 is located in the chloroplast. The analyses of the knockout (ossqd1) and knockdown (Ri1 and Ri2) mutants demonstrated reductions in Pi and total P concentrations, 32 Pi uptake rate, expression levels of Pi transporters and altered developmental responses of root traits, which were accentuated during Pi deficiency. The inhibitory effects of the OsSQD1 mutation were also evident in the development of reproductive tissue. Furthermore, OsSQD1 differently affects lipid composition under different Pi regime affects sulfur (S) homeostasis. Together, the study revealed that OsSQD1 affects Pi and S homeostasis, and lipid composition in response to Pi deprivation.

The ferroxidase LPR5 functions in the maintenance of phosphate homeostasis and is required for normal growth and development of rice.

Members of the Low Phosphate Root (LPR) family have been identified in rice (Oryza sativa) and expression analyses have been conducted. Here, we investigated the functions of one of the five members in rice, LPR5. qRT-PCR and promoter-GUS reporter analyses indicated that under Pi-sufficient conditions OsLPR5 was highly expressed in the roots, and specific expression occurred in the leaf collars and nodes, and its expression was increased under Pi-deficient conditions. In vitro analysis of the purified OsLPR5 protein showed that it exhibited ferroxidase activity. Overexpression of OsLPR5 triggered higher ferroxidase activity, and elevated concentrations of Fe(III) in the xylem sap and of total Fe in the roots and shoots. Transient expression of OsLPR5 in Nicotiana benthamiana provided evidence of its subcellular localization to the cell wall and endoplasmic reticulum. Knockout mutation in OsLPR5 by means of CRISPR-Cas9 resulted in adverse effects on Pi translocation, on the relative expression of Cis-NATOsPHO1;2, and on several morphological traits, including root development and yield potential. Our results indicate that ferroxidase-dependent OsLPR5 has both a broad-spectrum influence on growth and development in rice as well as affecting a subset of physiological and molecular traits that govern Pi homeostasis.

Interconversion and functional composites of metal-organic frameworks and hydrogen-bonded organic frameworks.

Metal-organic frameworks (MOFs), an emerging class of highly ordered crystalline porous materials, possess structural tunability, high specific surface area, well-defined pores, and diverse pore environments and morphologies, making them suitable for various potential applications. Moreover, hydrogen-bonded organic frameworks (HOFs), constructed from organic molecules with complementary hydrogen-bonding patterns, are rapidly evolving into a novel category of porous materials due to their facile mild preparation conditions, solution processability, easy regeneration capability, and excellent biocompatibility. These distinctive advantages have garnered significant attention across diverse fields. Considering the inherent binding affinity between MOFs and HOFs along with the fact that many MOF linkers can serve as building blocks for constructing HOFs, their combination holds promise in creating functional materials with enhanced performance. This feature paper provides an introduction to the interconversion between MOFs and HOFs followed by highlighting the emerging applications of MOF-HOF composites. Finally, we briefly discuss the current challenges associated with future perspectives on MOF-HOF composites.

Ecological restoration enhanced the stability of epiphytic microbial food webs of submerged macrophytes: Insights from predation characteristics of epiphytic predators.

The application of various submerged macrophytes for ecological restoration has gained increasing attention in urban lake ecosystems. The multitrophic microbial communities that colonized in various submerged macrophytes constitute microbial food webs through trophic cascade effects, which affect the biogeochemical cycles of the lake ecosystem and directly determine the effects of ecological restoration. Therefore, it is essential to reveal the diversity, composition, assembly processes, and stability of the microbial communities within epiphytic food webs of diverse submerged macrophytes under eutrophication and ecological restoration scenarios. In this study, we explored the epiphytic microbial food webs of Vallisneria natans and Hydrilla verticillata in both eutrophic and ecological restoration regions. The obtained results indicated that the two regions with different nutrient levels remarkably affected the diversity and composition of epiphytic multitrophic microbial communities of submerged macrophytes, among them, the community composition of epiphytic predators were more prone to change. Secondly, environmental filtering effects played a more important role in driving the community assembly of epiphytic predators than that of prey. Furthermore, the generality and intraguild predation of epiphytic predators were significantly improved within ecological restoration regions, which increased the stability of epiphytic microbial food webs. Additionally, compared with Hydrilla verticillata, the epiphytic microbial food webs of Vallisneria natans exhibited higher multitrophic diversity and higher network stability regardless of regions. Overall, this study focused on the role of the epiphytic microbial food webs of submerged macrophytes in ecological restoration and uncovered the potential of epiphytic predators to enhance the stability of microbial food webs, which may provide new insights into the development of ecological restoration strategies.

Effects of different emergent macrophytes on methane flux and rhizosphere microbial communities in wetlands.

Emergent macrophytes are of great importance for the structure and functioning of wetland ecosystems and play a significant role in environmental improvement, element cycling, and greenhouse gas (GHG) emissions. However, our understanding of how GHG fluxes differ among macrophyte species and its links with the microbial communities remain limited. In this study, we investigated the rhizosphere microbial communities (including total bacteria, methanotrophs, and methanogens) and the GHG fluxes associated with four emergent macrophytes-Phragmites australis, Thalia dealbata, Pontederia cordata, and Zizania latifolia-collected from Xuanwu Lake wetland, China. We observed the highest CH4 flux (FCH4) (9.35 ± 2.52 mg·m-2·h-1) from Z. latifolia zone, followed by P. australis, P. cordata, and T. dealbata zones (5.38 ± 1.63, 2.38 ± 2.91, and 2.02 ± 0.69 mg·m-2·h-1, respectively). Methanogenesis was methylotrophic at all sites, as the 13C-CH4 values were higher than -64 ‰ and the fractionation coefficients were lower than 1.055. We found a positive linear relationship between FCH4 and the methanogen community, in particular the relative abundances of Methanobacterium and Methanosarcina, indicating that the variations in FCH4 among the studied macrophyte-dominated zones might be attributed to the differences in rhizosphere microbial communities. The methane emissions in various macrophyte zones might be due to the higher capacity of methanogenesis compared to methane oxidation which was inhibited by nutrient-rich sediments. Our findings provide insights for selecting specific emergent macrophytes characterized by low FCH4 in wetland ecological restoration.

Luminescent Porous Organic Crystals for Adsorptive Separation of Toluene and Methylcyclohexane.

A butterfly-shaped phenothiazine derivative, PTTCN, was synthesized to obtain pure organic porous crystals for the highly efficient absorptive separation of toluene (Tol) and methylcyclohexane (Mcy). Due to the presence of three polar cyano groups and nonplanar conformation, these molecules self-assembled into a hydrogen-bonded organic framework (X-HOF-5) with distinct cavities capable of accommodating Tol molecules through multiple hydrogen-bonding interactions. Upon solvent removal via heating, the activated X-HOF-5 retained its cavity structure albeit with altered stacking arrangements, accompanied by a remarkable fluorescent color change from cyan to green. X-HOF-5a can undergo a phase transformation into X-HOF-5 upon reabsorption of Tol, while exhibiting no accommodation of Mcy due to the weak intermolecular interaction between PTTCN and Mcy. This suggests that the activated HOF material prefers Tol over Mcy. Moreover, X-HOF-5a may selectively accommodate Tol in a Tol/Mcy equimolar mixture, and the purity of Tol can reach 97% after release from the framework. Additionally, it is noteworthy that the HOF material exhibits recyclability without any discernible loss in performance.

A novel epithelial-mesenchymal transition-related lncRNA signature predicts prognosis and immune status in endometrioid endometrial cancer.

The pathogenesis and progression of endometrial cancer (EC) are associated with epithelial-mesenchymal transition (EMT) and long noncoding RNAs (lncRNAs). In the present study, we aimed to identify an EMT-related lncRNA signature and evaluate its prognostic value in EC. We obtained the expression profile of lncRNAs and clinical information of patients with endometrioid EC from The Cancer Genome Atlas database (N = 401). We identified a signature of 5 EMT-related lncRNAs and calculated the risk score of each patient. Next, we validated the independence of the prognostic value of the EMT-related lncRNA signature. Furthermore, we performed Gene Set Enrichment Analysis to identify potential molecular function and Kyoto Encyclopedia of Genes and Genomes pathways related to the EMT-related lncRNA signature. Tumor microenvironment analysis and immune checkpoint blockade (ICB) response prediction were also assessed. Survival analysis revealed that the high-risk group, based on the EMT-related lncRNA signature, had a poorer prognosis than the low-risk group in the training, testing, and entire sets. The predictive value of the EMT-related lncRNA signature was independent of age, The International Federation of Gynecology and Obstetrics stage, tumor grade, and body mass index. Time-dependent receiver operating characteristic curves also demonstrate the prognostic accuracy of this risk model. Gene Set Enrichment Analysis showed that cytokine-cytokine receptor interaction, glycolysis/gluconeogenesis, and IL-17 signaling pathway were significantly enriched. Furthermore, tumor microenvironment analysis indicated a significant negative correlation between the immune score and EMT-related lncRNA signature risks core, while the low-risk group was more likely to respond to ICB therapy than the high-risk group. A reliable EMT-related lncRNA signature of endometrioid EC was identified that could be utilized as an independent prognostic biomarker to predict patient survival outcomes and provide references for the option of ICB therapy.

Greater transmission capacities and small-world characteristics of bacterial communities in the above- than those in the below- ground niches of a typical submerged macrophyte, Vallisneria natans.

Leaves and roots of submerged macrophytes provide extended surfaces and stable internal tissues for distinct microorganisms to rest, but how these microorganisms interact with each other across different niches and ultimately drive the distribution through horizontal and vertical transmissions remains largely undetermined. Knowledge of the mechanisms of assemblage and transmission in aquatic macrophytes-associated microbial communities will help to better understanding their important roles in plant fitness and benefit ecological functions. Here, we conducted a microcosmic experiment based on in situ lake samples to investigate the bacterial community assemblage, transmission, and co-occurrence patterns in different niches of a typical submerged macrophyte, Vallisneria natans (V. natans), including seed endosphere, as well as environmental (water and bulk sediment), epiphytic (phyllosphere and rhizosphere), and endophytic (leaf and root endosphere) microhabitats of both leaves and roots representatives of the above- and below- ground niches (AGNs and BGNs), respectively. We found the bacterial communities colonized in epiphytic niches not only exhibited the highest diversity compared to adjacent environmental and endophytic niches, but also dominated the interactions between those bacterial members of neighboring niches in both AGNs and BGNs. The host plants promoted niche specificity at bacterial community-level, as confirmed by the proportion of bacterial specialists increased with plant proximity, especially in the BGNs. Furthermore, the bacterial taxa colonized in the AGNs exhibited higher horizontal and vertical transmission capacities than those in the BGNs, especially in the vertical transmission from seeds to leaves (41.38 %) than roots (0.42 %). Meanwhile, the bacterial co-occurrence network in AGNs was shown to have stronger small-world characteristics but weaker stability than those in the BGNs. Overall, this study cast new light on the plant microbiome in the aquatic environment, thus better promoting the potential development of strategies for breeding aquatic macrophyte holobiont with enhanced water purification and pollutant removal capabilities in the future.

A Multistimuli Responsive, Flexible Luminescent Framework and Its Applicability in Anticounterfeiting.

A linear distyrylanthracene derivative (DDATAn) with two diaminotriazine (DAT) groups acting as the hydrogen bond (H-bond) units was designed and synthesized in order to construct flexible organic porous crystals. H-bonds between the DAT moieties helped the molecules to construct a double interpenetrated two-dimensional layer, and the stacking between layers provided a H-bonded organic framework (X-HOF-3) with one-dimensional solvent channels. When X-HOF-3 was placed in contact with methanol, the fluorescent colors of the HOF exhibited an apparent bathochromic shift. More interestingly, the methanol-activated HOF was able to rapidly adsorb water from the air, which was accompanied by a change in fluorescent color from yellow to red. Under heating, water was released from the HOF and the fluorescent color returned to yellow. Water molecules in the pores were also able to be released after an applied mechanical force disrupted the ordered structure of the HOF. Based on these stimuli-responsive properties, these HOFs can be used as advanced functional materials in anticounterfeiting applications.

Testosterone elevation in ovarian adult granulosa cell tumor: A case report and review of the literature.

RATIONALE: Adult granulosa cell tumors (AGCT) mainly secret estrogen, but few androgens. It is rarer to have amenorrhea and hyperandrogenemia as clinical features. Here, we report a rare case of right side AGCTs with amenorrhea and hyperandrogenemia in a 19-year-old female. PATIENT CONCERNS: The 19-year-old patient was admitted to our hospital due to amenorrhea for more than 1 year, and discovery of pelvic mass for 4 months. The gynecological ultrasound and computed tomography (CT) cannot define the nature of the mass. Surprisingly, an elevation in testosterone levels was also measured. DIAGNOSIS AND INTERVENTIONS: The present patient underwent laparoscopic right salpingo-oophorectomy and partial omentectomy and biopsy of the peritoneum. OUTCOMES: After the surgery, the testosterone value was down to normal. The patient menstrual cramps on August 13, 2021. Her clitoris is smaller than the front. Up to August 1, 2022, there was no obvious sign of recurrence. LESSONS: Androgen-secreting AGCT is rare. We hope that this case can strengthen gynecologists' early diagnosis and treatment of this disease and improve the prognosis.

Cumulative ecosystem response to Hydraulic Engineering Infrastructure Projects in an arid basin.

Hydraulic Engineering Infrastructure Projects (HEIPs) typically show profound effects on hydrological systems and ecosystems. However, data restrictions have limited the exploration of the influences of compound HEIPs on ecosystems to a few studies. This study proposes a watershed-wide ecosystem assessment framework to investigate the impact of HEIPs in the Tarim River Headwaters-Hotan River Basin on the ecosystem of the arid zone. The framework includes a deep learning-meta cellular automata algorithm (DLMCAA) based on the spatiotemporal characteristics of HEIPs and hydro-meteorological and human activities. Moreover, the spatiotemporal relationships between compound HEIPs and ecosystem variances were quantified. The framework including DLMCAA showed a good performance in simulating landcover in 2020, with a Kappa coefficient of 0.89. Therefore, the DLMCAA could be used to simulate and predict ecosystem changes under the HEIPs, which suggested that the framework is effective and practical. An analysis of the spatiotemporal distribution of each ecosystem from 1980 to 2020 showed that the low shrub ecosystems changed most significantly (26.38 %) between 1980 and 2020. Also, the use of spatially driven hydrological project data from different ABC scenarios showed that ecosystems driven by HEIPs were more stable compared to those without HEIPs under future climate change. In particular, the DLMCAA indicated that compound HEIPs had a more positive impact on ecosystem oases in arid lands compared with that of single HEIPs. The results of this study can serve as a scientific reference for assessing the impact of HEIPs, as well as for understanding ecosystem changes and facilitating sustainable water resource management in the arid regions.

Overexpression of OsPHR3 improves growth traits and facilitates nitrogen use efficiency under low phosphate condition.

Phosphorus (P) and nitrogen (N) are both essential macronutrients for maintaining plant growth and development. In rice (Oryza sativa L.), OsPHR3 is one of the four paralogs of PHR1, which acts as a central regulator of phosphate (Pi) homeostasis, as well being involved in N homeostasis. However, the functions of OsPHR3 in N utilization under different Pi conditions have yet to be fully studied. In this study, we aimed to dissect the effect of OsPHR3-overexpression on N utilization under Pi deficient regimes. Biochemical, molecular and physiological assays were performed to determine the N-influx, translocation, and accumulation in OsPHR3-overexpressing rice lines, grown under Pi-sufficient and -deficient conditions, in both hydroponic and soil systems. Furthermore, important agronomic traits of these plants were also evaluated. The overexpression of OsPHR3 increased N uptake under Pi stress regimes. Increased N uptake also elevated total N concentrations in these plants by inducing N transporter genes expression. Furthermore, overexpression of OsPHR3 increased N use efficiency, 1000-grain weight and grain yield under different Pi conditions. We established new findings that OsPHR3-overexpression facilitates N utilization under Pi deficient conditions. This will help achieving higher yields by coordinating the utilization of N and P.

OsPDR2 mediates the regulation on the development response and maintenance of Pi homeostasis in rice.

Inorganic orthophosphate (Pi), a major form of essential macronutrient phosphorus (P), is available in rhizosphere for acquisition and assimilation by plants. However, the limited availability of Pi in soils affects the growth and development of plants. In Arabidopsis thaliana (Arabidopsis), Phosphate Deficiency Response2 (AtPDR2), interacts genetically with Low Phosphate Root1 (AtLPR1) in the endoplasmic reticulum (ER) and plays a key role in the inhibition of primary root growth (PRG) during Pi deficiency. However, the role of OsPDR2, the homolog of AtPDR2, either in roots response to Pi deficiency and/or in growth and development has not been elucidated as yet. Therefore, qRT-PCR was employed to determine the spatiotemporal effects and the availability of Pi on the expression of OsPDR2. OsPDR2 showed variable levels of relative expression pattern in vegetative and/or reproductive tissues analyzed at different stages of growth and development (5-17 weeks). Transient expression analysis revealed its subcellular localization to the ER. Further, the reverse genetics approach was employed for determining the function of OsPDR2 by generating RNAi lines (Ri2, Ri9, and Ri18). The study revealed significant inhibitory effects of RNAi-mediated suppression of OsPDR2 on the development of root, male reproductive traits, and yield. Moreover, 32P isotope labeling and split-root experiments under different Pi regime with RNAi lines revealed the function of OsPDR2 in regulating homeostasis of Pi.

Lack of interferon response in animals to naked siRNAs.

RNA interference (RNAi) is rapidly becoming the method of choice for the elucidation of gene function and the identification of drug targets. As with other oligonucleotide-based strategies, RNAi is envisioned to ultimately be useful as a human therapeutic. Unlike previous nucleic acid therapeutics, small interfering RNAs have the potential to elicit immune responses via interactions with Toll-like receptor 3 and trigger interferon responses like long, double-stranded RNA and its analogs, such as poly(I:C). Recently, the safety of siRNAs has been questioned because they have been shown to trigger an interferon response in cultured cells. We show here that it is possible to administer naked, synthetic siRNAs to mice and downregulate an endogenous or exogenous target without inducing an interferon response.

Selective usage of D-Type cyclins by Ewing's tumors and rhabdomyosarcomas.

The genetic mechanisms that control proliferation of childhood musculoskeletal malignancies, notably Ewing's tumor (ET) and rhabdomyosarcoma (RMS), remain largely unknown. Most human cancers appear to overexpress at least one of the G1 cyclins (cyclins D1, D2, D3, E1, and E2) to bypass normal regulation of cell cycle G1 progression. We compared the gene expression profiles of 7 ET and 13 RMS primary tumor samples and found overexpression of cyclin D1 in all 7 ET samples. In contrast, RMS samples expressed higher levels of cyclin D2, cyclin D3, and cyclin E1. This was confirmed by quantitative reverse transcription-polymerase chain reaction and Western blot. The relative roles of RAS-extracellular signal-regulated kinase 1/2 and phosphatidylinositol 3'-kinase (PI3K)-AKT pathways in the regulation of D-type cyclin expression in these tumors were then assessed. Inhibition of either pathway reduced expression of cyclins D1, D2, and D3 in RMS lines, whereas only PI3K inhibitors blocked cyclin D1, D2, and D3 expression in ET lines. Furthermore, PI3K-AKT appeared to regulate D-type cyclin transcription in RMS lines through FKHR and FKHRL1. Finally, the role of the ET-associated EWS-FLI1 fusion gene in regulating D cyclin expression was studied. Inhibition of EWS-FLI1 expression in the TC71 ET line decreased cyclin D1 levels but increased cyclin D3 levels. In contrast, induction of EWS-FLI1 expression in the RD RMS cell line increased cyclin D1 expression but decreased cyclin D3 expression. Our results demonstrate distinct regulation of D-type cyclins in ET and RMS and indicate that EWS-FLI1 can modulate the expression of D-type cyclins independent of cellular backgrounds.