Reducing phenanthrene uptake and translocation, and accumulation in the seeds by overexpressing OsNRT2.3b in rice.

The uptake and accumulation of polycyclic aromatic hydrocarbons (PAHs) in crops have gained much attention due to their toxicity to humans. Nitrogen (N) is an essential element for plant growth and has also been implicated in the acquisition and acropetal translocation of PAHs. OsNRT2.3b encodes a nitrate (NO3-) transporter that is involved in the acquisition and mobilization of N in rice. Here, we investigated whether overexpression of OsNRT2.3b would exert any mitigating influence on the uptake and translocation of phenanthrene (Phe, a model PAH) in transgenic rice (Oryza sativa). The wild-type seedlings exhibited a reduction in plant height, primary root length, and shoot biomass when grown hydroponically in a medium supplemented with Phe. Acquisition of Phe by the roots and its subsequent translocation to shoots increased concomitantly with an increase in Phe concentration in the medium and duration of the treatment. OsNRT2.3b-overexpressing lines (Ox-6 and Ox-8) were generated independently. Compared with the wild-type, the concentration of Phe in Ox-6 and Ox-8 were significantly lower in the roots (47%-54%) and shoots (22%-31%) grown hydroponically with Phe (1 mg/L). Further, the wild-type and Ox lines were grown to maturity in a pot soil under Phe conditions and the concentrations of Phe and total N were assayed in the culms and flag leaves. Compared with the wild-type, in Ox lines the concentration of total N significantly increased in the culms (288%-366%) and flag leaves (12%-25%), while that of Phe significantly reduced in the culms (25%-28%) and flag leaves (18%-21%). The results revealed an antagonistic correlation between the concentration of total N and Phe. The concentration of Phe was also significantly lower (29%-38%) in the seeds of Ox lines than the wild-type. The study highlighted the efficacy of overexpressing OsNRT2.3b in mitigating the Phe toxicity by attenuating its acquisition, mobilization, and allocation to the seeds.

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.

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.

Fine particulate matter exposure induces cell cycle arrest and inhibits migration and invasion of human extravillous trophoblast, as determined by an iTRAQ-based quantitative proteomics strategy.

Long-term exposure to fine particulate matter (PM2.5) may cause adverse pregnancy outcomes but the mechanisms are not clear. Our research confirms that PM2.5 induced DNA damage, and inhibited cell proliferation in HTR-8/SVneo cells, presenting in a dose- and time-dependent manners. Using quantitative proteomics, the 182 and 486 differentially expressed proteins in cells treated with 120µgml-1 PM2.5 for 24 and 48h were involved in many critical biological processes, including of cell proliferation, response to DNA damage, regulation of small GTPase mediated signal transduction, and etc. Further validation indicated that PM2.5 blocked the cell cycle at the G2/M phase through activation of the ATR-Cyclin E1/Cdk6 pathway, and it reduced the migration and invasion by upregulating TIMP1 and TIMP2 expression and downregulating Collagen I expression. Our findings were consistent with the observed effects of PM2.5 on cell cycle arrest and inhibition of migration and invasion in human extravillous trophoblast.

[Screening and identification of differential expression genes related to stress fracture by cDNA microarray assay].

OBJECTIVE: To evaluate the differentially expressed genes between the Stress fracture (SF) cases and controls. METHODS: Total RNA was extracted and purified from peripheral blood sample of 3 SF cases and 3 controls who conducted a 1:1 matched case-control study, then used for Human Genome Array analysis. The hybridization data were analyzed using SAM software. Parts of these genes were analyzed and identified by real-time PCR. RESULTS: Upregulated and downregulated genes were 22 and 1, respectively. Thus the highest ratio and most significant cytokine was tumor necrosis factor receptor superfamily, member 10c (TNFRSF10C). The result of real-time PCR shows that TNFRSF10C was over-expressed in 3 cases and low-expressed in 1 case. CONCLUSION: Obvious difference exists in gene expression between SF cases and controls, showing there may be a lot of genes involving in the occurrence and development of SF. Meanwhile, the identification of the specific genes is helpful for biomechanics study, early diagnosis and screening of SF.