Comparative transcriptome and DNA methylation analysis in temperature-sensitive genic male sterile wheat BS366.

BACKGROUND: Known as the prerequisite component for the heterosis breeding system, the male sterile line determines the hybrid yield and seed purity. Therefore, a deep understanding of the mechanism and gene network that leads to male sterility is crucial. BS366, a temperature-sensitive genic male sterile (TGMS) line, is male sterile under cold conditions (12 °C with 12 h of daylight) but fertile under normal temperature (20 °C with 12 h of daylight). RESULTS: During meiosis, BS366 was defective in forming tetrads and dyads due to the abnormal cell plate. During pollen development, unusual vacuolated pollen that could not accumulate starch grains at the binucleate stage was also observed. Transcriptome analysis revealed that genes involved in the meiotic process, such as sister chromatid segregation and microtubule-based movement, were repressed, while genes involved in DNA and histone methylation were induced in BS366 under cold conditions. MethylRAD was used for reduced DNA methylation sequencing of BS366 spikes under both cold and control conditions. The differentially methylated sites (DMSs) located in the gene region were mainly involved in carbohydrate and fatty acid metabolism, lipid metabolism, and transport. Differentially expressed and methylated genes were mainly involved in cell division. CONCLUSIONS: These results indicated that the methylation of genes involved in carbon metabolism or fatty acid metabolism might contribute to male sterility in BS366 spikes, providing novel insight into the molecular mechanism of wheat male sterility.

Genome-wide identification and transcriptional characterization of DNA methyltransferases conferring temperature-sensitive male sterility in wheat.

BACKGROUND: DNA methyltransferase (DMT) genes contribute to plant stress responses and development by de novo establishment and subsequent maintenance of DNA methylation during replication. The photoperiod and/or temperature-sensitive genic male sterile (P/TGMS) lines play an important role in hybrid seed production of wheat. However, only a few studies have reported on the effect of DMT genes on temperature-sensitive male sterility of wheat. Although DMT genes have been investigated in some plant species, the identification and analysis of DMT genes in wheat (Triticum aestivum L.) based on genome-wide levels have not been reported. RESULTS: In this study, a detailed overview of phylogeny of 52 wheat DMT (TaDMT) genes was presented. Homoeolog retention for TaDMT genes was significantly above the average retention rate for whole-wheat genes, indicating the functional importance of many DMT homoeologs. We found that the strikingly high number of TaDMT genes resulted mainly from the significant expansion of the TaDRM subfamily. Intriguingly, all 5 paralogs belonged to the wheat DRM subfamily, and we speculated that tandem duplications might play a crucial role in the TaDRM subfamily expansion. Through the transcriptional analysis of TaDMT genes in a TGMS line BS366 and its hybrids with the other six fertile lines under sterile and fertile conditions, we concluded that TaCMT-D2, TaMET1-B1, and TaDRM-U6 might be involved in male sterility in BS366. Furthermore, a correlation analysis showed that TaMET1-B1 might negatively regulate the expression of TaRAFTIN1A, an important gene for pollen development, so we speculated regarding an epigenetic regulatory mechanism underlying the male sterility of BS366 via the interaction between TaMET1-B1 and TaRAFTIN1A. CONCLUSIONS: Our findings presented a detailed phylogenic overview of the DMT genes and could provide novel insights into the effects of DMT genes on TGMS wheat.

Transcriptome analysis of wheat seedling and spike tissues in the hybrid Jingmai 8 uncovered genes involved in heterosis.

MAIN CONCLUSION: Transcriptome analysis was carried out for wheat seedlings and spikes from hybrid Jingmai 8 and both inbred lines to unravel mechanisms underlying heterosis. Heterosis, known as one of the most successful strategies for increasing crop yield, has been widely exploited in plant breeding systems. Despite its great importance, the molecular mechanism underlying heterosis remains elusive. In the present study, RNA sequencing (RNA-seq) was performed on the seedling and spike tissues of the wheat (Triticum aestivum) hybrid Jingmai 8 (JM8) and its homozygous parents to unravel the underlying mechanisms of wheat heterosis. In total, 1686 and 2334 genes were identified as differentially expressed genes (DEGs) between the hybrid and the two inbred lines in seedling and spike tissues, respectively. Gene Ontology analysis revealed that DEGs from seedling tissues were significantly enriched in processes involved in photosynthesis and carbon fixation, and the majority of these DEGs expressed at a higher level in JM8 compared to both inbred lines. In addition, cell wall biogenesis and protein biosynthesis-related pathways were also significantly represented. These results confirmed that a combination of different pathways could contribute to heterosis. The DEGs between the hybrid and the two inbred progenitors from the spike tissues were significantly enriched in biological processes related to transcription, RNA biosynthesis and molecular function categories related to transcription factor activities. Furthermore, transcription factors such as NAC, ERF, and TIF-IIA were highly expressed in the hybrid JM8. These results may provide valuable insights into the molecular mechanisms underlying wheat heterosis.

Improving the simultaneous removal of chemical oxygen demand and terephthalic acid in a cross-flow aerobic sludge reactor by using response surface methodology.

Central composite design and response surface methodology (RSM) were implemented to optimize the operational parameters for a cross-flow aerobic sludge reactor (CFASR) in remedying mixed printing and dyeing wastewater (MPDW). The individual and interactive effects of three variables, hydraulic retention time (HRT), pH and sludge loading rate (SLR), on chemical oxygen demand (COD) and terephthalic acid (TA) removal rates were evaluated. For HRT of 15.3-19.8 hours, pH of 7.2-8.1 and SLR of 0.4-0.6 kg chemical oxygen demand (COD) per kg mixed liquor suspended solids per day, COD and TA removal rates of the CFASR exceeded 85% and 90%, respectively. The check experiment revealed that the effluent from the optimized CFASR was stable below the limitation of 100 mg COD/L and the TA concentration decreased by 6.0% compared to the usual CFASR. The results verified that the RSM was useful for optimizing the operation parameters of the CFASR in remedying MPDW.

Transient wave propagation in the ring stiffened laminated composite cylindrical shells using the method of reverberation ray matrix.

The method of reverberation ray matrix is extended to investigate the transient wave propagation and early short time transient responses of the ring stiffened laminated composite cylindrical shells subjected to impact loads. The ring stiffened laminated cylindrical shells are modeled as the continuous coupling systems between the vibration of the un-stiffened laminated cylindrical shell and the motion of the curved beams. The dynamic models of the laminated cylindrical shell and curved beams in the Laplace phase space are established on the basis of the first order shear deformation theory. The reverberation ray matrix can be determined by the boundary and continuous conditions of the ring stiffened laminated cylindrical shell. Using the fast Fourier transform algorithm, the dynamic responses of the ring stiffened laminated cylindrical shells can be computed. Through the numerical simulations, it can be seen that the early short time transient accelerations of the ring stiffened laminated cylindrical shells under impact loads are very large, while the early short time transient shear strains and displacements are very small. Furthermore, the influences of the ring stiffener number and impact load types on the early short time transient responses of the ring stiffened laminated cylindrical shells are also investigated.

Feasibility and simulation model of a pilot scale membrane bioreactor for wastewater treatment and reuse from Chinese traditional medicine.

The lack and pollution of water resource make wastewater reuse necessary. The pilot scale long-term tests for submerged membrane bioreactor were conducted to treat the effluents of anaerobic or aerobic treatment process for the high-strength Chinese traditional medicine wastewater. This article was focused on the feasibility of the wastewater treatment and reuse at shorter hydraulic retention time (HRT) of 5.0, 3.2 and 2.13 h. MLSS growth, membrane flux, vacuum values and chemical cleaning periods were also investigated. The experimental results of treating two-phase anaerobic treatment effluent demonstrated that the CODfilt was less than 100 mg/L when the influent COD was between 500-10000 mg/L at HRT of 5.0 h, which could satisfy the normal discharged standard in China. The experimental results to treat cross flow aerobic reactor effluent demonstrated that the average value of CODfilt was 17.28 mg/L when the average value of influent COD was 192.84 mg/L at HRT of 2.13 h during 106 d, which could completely meet the normal standard for water reuse. The maximum MLSS and MLVSS reached 24000 and 14500 mg/L at HRT of 3.2 h respectively. Membrane flux had maximal resume degrees of 94.7% at vacuum value of 0.02 MPa after cleaning. Chemical cleaning periods of membrane module were 150 d. A simulation model of operational parameters was also established based on the theory of back propagation neural network and linear regression of traditional mathematical model. The simulation model showed that the optimum operational parameters were suggested as follows: HRT was 5.0 h, SRT was 100 d, the range of COD loading rate was between 10.664-20.451 kg/(m3xd), the range of MLSS was between 7543-13694 mg/L.

Performance and model of a novel multi-sparger multi-stage airlift loop membrane bioreactor to treat high-strength 7-ACA pharmaceutical wastewater: effect of hydraulic retention time, temperature and pH.

In this study, three novel multi-sparger multi-stage airlift loop membrane bioreactors (Ms(2)ALMBRs) were set up in parallel for treating synthetic high-strength 7-ACA pharmaceutical wastewater under different HRTs, temperatures and pHs, respectively. During the 200-day operating time, average COD removal efficiencies were 94.96%, 96.05% and 93.9%. While average 7-ACA removal efficiencies were 66.44%, 59.04% and 59.60%, respectively. The optimal conditions were 10h, 15-35°C and 7-9 for HRT, temperature and pH, respectively. Moreover, the sludge characteristics and microorganism drug-resistances were explored. Results showed that different temperatures and pHs influenced contaminant removals by affecting MLSS concentration and ß-lactamase activity significantly. In addition, mathematical statistical models, built on the polynomial and linear regression techniques, were developed for exploring the inner relationships between HRT, temperature and pH changes and MLSS concentrations, ß-lactamase activities and contaminant removals of the Ms(2)ALMBR system.

The inhibitory effects of NKX3.1 on IGF-1R expression and its signalling pathway in human prostatic carcinoma PC3 cells.

NKX3.1, which is a prostate-specific homeobox gene, plays an important role in prostate cancer and usually functions as a tumour suppressor gene. In this study, we investigated the inhibitory effect of NKX3.1 on insulin-like growth factor (IGF)-1R expression and its downstream signalling pathway in PC3 cells. PC3 cells were stably transfected with NKX3.1 expression plasmid (pcDNA3.1-NKX3.1) or vector plasmid (pcDNA3.1+). The IGF-IR mRNA and protein expression levels were assessed in PC3-NKX3.1 transfectants by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. The expression and activation of IGF-1/IGF-1R downstream signalling targets were examined by Western blotting and luciferase reporter assay. The cells were subsequently treated with relevant concentrations of IGF-1. The effect of IGF-1 on cell growth was examined by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide (MTT) assay and flow cytometry analysis. A significant suppression of IGF-1R mRNA and protein expression was observed after forced expression of NKX3.1 in PC3 cells. Correspondingly, the forced expression of NKX3.1 decreased IGF-1-induced phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (AKT) and activation of the Elk-1 transcription factor and downregulated the expression of the downstream target genes c-fos and cyclin D1. Furthermore, the forced expression of NKX3.1 inhibited IGF-1-induced cell growth. In conclusion, NKX3.1 could downregulate IGF-1R expression and could inhibit IGF-1R-mediated mitogen-activated protein kinase (MAPK)/ERK and AKT signalling pathways, which might partially leads to the inhibition of IGF-1-induced cell growth. This study provides new insights into the molecular mechanisms that NKX3.1 exerts against prostate cancer and ultimately expands the scope of alternative approaches in advanced prostate cancer therapy.

Improving the simultaneous removal efficiency of COD and color in a combined HABMR-CFASR system based MPDW. Part 1: optimization of operational parameters for HABMR by using response surface methodology.

The aim of this study was to implement central-composite design (CCD) and response surface methodology (RSM) to optimize the operational parameters for hybrid anaerobic baffled microbial reactor (HABMR) remedying mixed printing and dyeing wastewater (MPDW). The individual and interactive effects of three variables, hydraulic retention time (HRT), pH, sludge loading rate (SLR) on the COD and color removal rates were evaluated. In the area of HRT: 12.5-13.9 h, pH: 9.0-9.5 and SLR: 0.27-0.33 kg COD/(kg MLVSSd), COD and color removal rates of HABMR exceeded 40% and 60%, simultaneously. The check experiment revealed that the amount of COD and color in the effluent could be decreased by 9.97% and 10.12% compared to the usual operating conditions, respectively. The results verified that the RSM was useful for optimizing the operational parameters of HABMR in treating MPDW.

Improving the efficiencies of simultaneous organic substance and nitrogen removal in a multi-stage loop membrane bioreactor-based PWWTP using an on-line Knowledge-Based Expert System.

The results of the use of an expert system (ES) to control a novel multi-stage loop membrane bioreactor (MLMBR) for the simultaneous removal of organic substances and nutrients are reported. The study was conducted at a bench-scale plant for the purpose of meeting new discharge standards (GB21904-2008) for the treatment of chemical synthesis-based pharmaceutical wastewater (1200-9600 mg/L COD, 500-2500 mg/L BOD5, 50-200 mg/L NH4+-N and 105-400 mg/L TN in the influent water) by developing a distributed control system. The system allows various expert operational approaches to be deployed with the goal of minimizing organic substances and nitrogen levels in the outlet while using the minimum amount of energy. The proposed distributed control system, which is supervised by a Knowledge-Based Expert System (KBES) constructed with G2 (a tool for expert system development) and a back propagation BP artificial neural network, permits the on-line implementation of every operating strategy of the experimental system. A support vector machine (SVM) is applied to achieve pattern recognition. A set of experiments involving variable sludge retention time (SRT), hydraulic retention time (HRT) and dissolved oxygen (DO) was carried out. Using the proposed system, the amounts of COD, TN and NH4+-N in the effluent decreased by 55%, 62% and 38%, respectively, compared to the usual operating conditions. These improvements were achieved with little energy cost because the performance of the treatment plant was optimized using operating rules implemented in real time.

[Full-scale two-phase anaerobic process treating traditional Chinese medicine wastewater].

A navel two-phase anaerobic-aerobic treatment technology was developed to treat high concentration and refractory organic wastewater discharge from Harbin Second Traditional Chinese Medicine Factory. The function of two-phase anaerobic process in the whole bio-treatment system was analyzed. Now the status in bio-technique of treating wastewater from traditional Chinese medicine production was that all the influent COD concentration was under 5 000 mg/L and its quality was easy to be bio-treated. The full-scale experiment has changed this status. Full-scale experiment results showed that when influent COD concentration was at 7 000 - 40 000 mg/L and its BOD5 /COD was lower than 0.2, the volume-loading rate and COD removal rate of acidogenic phase, a continuous-flow stirred tank reactor, could reach 20- 30 kg/(m3 x d) and 47.1% respectively. At the same time, it only took the mathanogenic phase 27 days to set-up. The volume-loading rate and COD removal rate of mathanogenic phase, an up-flow anaerobic sludge blanket-anaerobic filter reactor, could reach 6.0 - 7.0 kg/(m3 d) and 94.06% respectively. It was also demonstrated the COD removal rate of two-phase anaerobic process was over than 93.0%, which offer endurance for the effluent from the whole treatment system reaching relative standard performed in China.