Identification of Key Lipogenesis Stages and Proteins Involved in Walnut Kernel Development.

Walnuts are abundant in oil content, especially for polyunsaturated fatty acids, but the understanding of their formation is limited. We collected walnut (Juglans regia L.) kernels at 60, 74, 88, 102, 116, 130, and 144 days after pollination (designated S1-S7). The ultrastructure and accumulation of oil bodies (OBs) were observed using transmission electron microscopy (TEM), and the oil content, fatty acid composition, and proteomic changes in walnut kernels were determined. The oil content and OB accumulation increased during the development and rose sharply from S1 to S3 stages, which are considered the key lipogenesis stage. A total of 5442 proteins were identified and determined as differentially expressed proteins (DEPs) using label-free proteomic analysis. Fatty acid desaturases (FAD) 2, FAD3, oleosin, and caleosin were essential and upregulated from the S1 to S3 stages. Furthermore, the highly expressed oleosin gene JrOLE14.7 from walnuts was cloned and overexpressed in transgenic Brassica napus. The overexpression of JrOLE14.7 increased the oil content, diameter, hundred weight of seeds and changed the fatty acid composition and OB size of Brassica napus seeds. These findings provide insights into the molecular mechanism of oil biosynthesis and the basis for the genetic improvement of walnuts.

Comprehensive Analyses of Simple Sequence Repeat (SSR) in Bamboo Genomes and Development of SSR Markers with Peroxidase Genes.

Simple sequence repeats (SSRs) are one of the most important molecular markers, which are widespread in plants. Bamboos are important forest resources worldwide. Here, the comprehensive identification and comparative analysis of SSRs were performed in three woody and two herbaceous bamboo species. Altogether 567,175 perfect SSRs and 71,141 compound SSRs were identified from 5737.8 Mb genome sequences of five bamboo species. Di-nucleotide SSRs were the most predominant type, with an average of ~50,152.2 per species. Most SSRs were located in intergenic regions, while those located in genic regions were relatively less. Moreover, the results of annotation distribution indicated that terms with P450, peroxidase and ATP-binding cassette transporter related to lignin biosynthesis might play important roles in woody and herbaceous bamboos under the mediation of SSRs. Furthermore, the peroxidase gene family consisted of a large number of genes containing SSRs was selected for the evolutionary relationship analysis and SSR markers development. Fifteen SSR markers derived from peroxidase family genes of Phyllostachys edulis were identified as polymorphic in 34 accessions belonging to seven genera in Bambusoideae. These results provided a comprehensive insight of SSR markers into bamboo genomes, which would facilitate bamboo research related to comparative genomics, evolution and marker-assisted selection.

An Integrated Regulatory Network of mRNAs, microRNAs, and lncRNAs Involved in Nitrogen Metabolism of Moso Bamboo.

Nitrogen is a key macronutrient essential for plant growth and development, and its availability has a strong influence on biological processes. Nitrogen fertilizer has been widely applied in bamboo forests in recent decades; however, the mechanism of nitrogen metabolism in bamboo is not fully elucidated. Here, we characterized the morphological, physiological, and transcriptome changes of moso bamboo in response to different schemes for nitrogen addition to illuminate the regulation mechanism of nitrogen metabolism. The appropriate addition of nitrogen improved the chlorophyll content and Pn (net photosynthetic rate) of leaves, the nitrogen and ammonium contents of the seedling roots, the biomass of the whole seedling, the number of lateral roots, and the activity of enzymes involved in nitrogen metabolism in the roots. Based on the whole transcriptome data of the roots, a total of 8,632 differentially expressed mRNAs (DEGs) were identified under different nitrogen additions, such as 52 nitrate transporter genes, 6 nitrate reductase genes, 2 nitrite reductase genes, 2 glutamine synthase genes, 2 glutamate synthase genes (GOGAT), 3 glutamate dehydrogenase genes, and 431 TFs belonging to 23 families. Meanwhile, 123 differentially expressed miRNAs (DEMs) and 396 differentially expressed lncRNAs (DELs) were characterized as nitrogen responsive, respectively. Furthermore, 94 DEM-DEG pairs and 23 DEL-DEG pairs involved in nitrogen metabolism were identified. Finally, a predicted regulatory network of nitrogen metabolism was initially constructed, which included 17 nitrogen metabolic pathway genes, 15 TFs, 4 miRNAs, and 10 lncRNAs by conjoint analysis of DEGs, DEMs, and DELs and their regulatory relationships, which was supported by RNA-seq data and qPCR results. The lncRNA-miRNA-mRNA network provides new insights into the regulation mechanism of nitrogen metabolism in bamboo, which facilitates further genetic improvement for bamboo to adapt to the fluctuating nitrogen environment.

Identification and Expression Analyses of Invertase Genes in Moso Bamboo Reveal Their Potential Drought Stress Functions.

Invertases (INVs) can irreversibly hydrolyze sucrose into fructose and glucose, which play principal roles in carbon metabolism and responses to various stresses in plants. However, little is known about the INV family in bamboos, especially their potential function in drought stress. In this study, 29 PeINVs were identified in moso bamboo (Phyllostachys edulis). They were clustered into alkaline/neutral invertase (NINV) and acid invertase (AINV) groups based on the gene structures, conserved motifs, and phylogenetic analysis results. The collinearity analysis showed nine segmental duplication pairs within PeINVs, and 25 pairs were detected between PeINVs and OsINVs. PeINVs may have undergone strong purification selection during evolution, and a variety of stress and phytohormone-related regulatory elements were found in the promoters of PeINVs. The tissue-specific expression analysis showed that PeINVs were differentially expressed in various moso bamboo tissues, which suggested that they showed functional diversity. Both the RNA-seq and quantitative real-time PCR results indicated that four PeINVs were significantly upregulated under drought stress. Co-expression network and Pearson's correlation coefficient analyses showed that these PeINVs co-expressed positively with sugar and water transport genes (SWTGs), and the changes were consistent with sugar content. Overall, we speculate that the identified PeINVs are spatiotemporally expressed, which enables them to participate in moso bamboo growth and development. Furthermore, PeINVs, together with SWTGs, also seem to play vital roles in the response to drought stress. These results provide a comprehensive information resource for PeINVs, which will facilitate further study of the molecular mechanism underlying PeINVs involvement in the response to drought stress in moso bamboo.

3D-QSAR-aided toxicity assessment of synthetic musks and their transformation by-products.

Synthetic musks (SMs) are fragrance additives widely used in personal care products. SMs and their transformation by-products may reach the environment even after wastewater treatment, resulting in ecological and health concerns. The identification and toxicity assessment of SM by-products generated from different chemical and biological treatment processes have been rarely studied. This study established a 3D-QSAR model based on SMs' molecular structures (independent variable) and their lethal concentration (LC50) of mysid (dependent variable). The developed model was further used to predict the LC50 of SMs transformation by-products. Fifty-eight by-products of six common SMs (i.e., galaxolide (HHCB), tonalide (AHTN), phantolide (PHAN), traseolide (TRASE), celestolide (ADBI), and musk ketone (MK)) generated from biodegradation, photodegradation, advanced oxidation, and chlorination were identified through literature review and lab experiment as the model inputs. Predicted LC50 results indicated that the toxicity of 40% chlorination by-products is higher than their precursors. Biodegradation is an effective method to treat AHTN. The advanced oxidation may be the best way to treat HHCB. This is the first study on biotoxicity of SM transformation by-products predicted by the 3D-QSAR model. The research outputs helped to provide valuable reference data and guidance to improve management of SMs and other emerging contaminants.

Extraction of methylmercury and ethylmercury from aqueous solution using surface sulfhydryl-functionalized magnetic mesoporous silica nanoparticles.

Surface sulfhydryl-functionalized magnetic mesoporous silica nanoparticles were prepared, aiming to extract trace alkylmercury from aqueous solution. The prepared nanoparticles were characterized by TEM, ED, EDX, DLS, FTIR, and SERS. Compare with that the non-sulfhydryl-functionalized Fe3O4@SiO2 exhibited almost no affinity for CH3Hg(+) and CH3CH2Hg(+); the sulfhydryl-functionalized Fe3O4@SiO2 exhibited high adsorption affinity for them, resulting from chelating interaction by surface sulfhydryl group, and the adsorption was not significantly impacted by pH within the range of 3.5-9.0 or coexisting metal ions. The monolayer adsorption on surface of Fe3O4@SiO2-RSH could reach equilibrium in 2 min. Moreover, the CH3Hg(+) and CH3CH2Hg(+) adsorbed on Fe3O4@SiO2-RSH could be quickly separated from the matrix in a magnetic field and desorbed easily by acetonitrile and l-cysteine aqueous solution or HCl solution, and the recoveries were more than 80%. Findings of the present work highlight the potential for using Fe3O4@SiO2-RSH magnetic nanoparticles as effective and reusable adsorbents for extraction of ultra trace alkylmercury from environmental water samples.

[Efficiency of micro-pressurized fluidized hybrid biological reactor for treating phenolic wastwater].

A dynamic experiment was conducted continuously to study the efficiency of the new micro-pressurized fluidized hybrid biological reactor (MP-FHBR) for treating artificial phenolic wastewater. The experimental results showed that the COD volumetric loading and phenolic volumetric loading of MP-FHBR could reach 4.86 kg/(m3 x d) and 1.96 kg/(m3 x d) respectively; effluent COD was lower than 200 mg/L; phenol concentrations of effluent were lower than 1 mg/L; removal rates of COD and phenol reached 90% and 99% above respectively. When volumetric loading of phenol in MP-FHBR reached 2.04 kg/(m3 x d), phenol will accumulate gradually in the reactor along with the reactor running, and specific TTC-dehydrogenase activity of microbe in the reactor declines. When phenol concentration of influent was 800 mg/L, the optimum HRT was 9-10 h. Because of the influence of hydraulic loading on the settling performance of sedimentation tank of the reactor, MP-FHBR was superior to treating higher-concentration phenolic wastewater.