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The anammox bacteria (AnAOB) have been suggested to perform dissimilatory Fe0/Fe2+-dependent nitrate reduction to ammonium (DNRA), with nitrite as intermediate. However, evidence at genetic and tra...

Weighted gene co-expression network analysis (WGCNA) is a research method in systematic biology. It is widely used to identify gene modules related to target traits in multi-sample transcriptome data. In order to further explore the molecular mechanism of potato response to drought stress, two cultivars (C16: CIP 397077.16 and C119: CIP 398098.119) introduced from the International Potato Center were used as experimental materials, and five drought stress gradients were treated for 0 h, 2 h, 6 h, 12 h, and 24 h, with the untreated material as a control. A total of 30 samples of root system were used for transcriptome sequencing, with three biological replicates. Based on the above transcriptome data, we constructed a co-expression network of weighted genes associated with stress-resistant physiological traits by WGCNA, and obtained 15 gene co-expression modules closely related to root drought resistance. In addition, a number of hub genes involved in drought stress regulation pathways were discovered from the four modules with the highest correlation with target traits. These results provide clues for further study on the molecular genetic mechanisms of potato root drought resistance.

Obesity is becoming a global problem. Research into the detailed mechanism of adipocyte development is crucial for the treatment of excess fat. Zinc finger protein 217 plays roles in adipogenesis. However, the underlying mechanism remains unclear. Here, we demonstrated that ZFP217 knockdown prevented the mitotic clonal expansion process and caused adipogenesis inhibition. Depletion of ZFP217 increased the expression of the m6A methyltransferase METTL3, which upregulated the m6A level of cyclin D1 mRNA. METTL3 knockdown rescued the siZFP217-inhibited MCE and promoted CCND1 expression. YTH domain family 2 recognized and degraded the methylated CCND1 mRNA, leading to the downregulation of CCND1. Consequently, cell-cycle progression was blocked, and adipogenesis was inhibited. YTHDF2 knockdown relieved siZFP217-inhibited adipocyte differentiation. These findings reveal that ZFP217 knockdown-induced adipogenesis inhibition was caused by CCND1, which was mediated by METTL3 and YTHDF2 in an m6A-dependent manner. We have provided novel insight into the underlying molecular mechanisms by which m6A methylation is involved in the ZFP217 regulation of adipogenesis.

N6-methyladenosine (m6A), the most abundant internal mRNA modification in eukaryotes, plays a vital role in regulating adipogenesis. However, its underlying mechanism remains largely unknown. Here, we reveal that deletion of m6A demethylase FTO in porcine and mouse preadipocytes inhibits adipogenesis through JAK2-STAT3-C/EBPβ signaling. Mechanistically, FTO deficiency suppresses JAK2 expression and STAT3 phosphorylation, leading to attenuated transcription of C/EBPβ, which is essential for the early stage of adipocyte differentiation. Using dual-luciferase assay, we validate that knockdown of FTO reduces expression of JAK2 in an m6A-dependent manner. Furthermore, we find that m6A "reader" protein YTHDF2 directly targets m6A-modified transcripts of JAK2 and accelerates mRNA decay, which results in decreased JAK2 expression and inactivated JAK2-STAT3-C/EBPβ signaling, thereby inhibiting adipogenesis. Collectively, our results provide a novel insight into the molecular mechanism of m6A methylation in post-transcriptional regulation of JAK2-STAT3-C/EBPβ signaling axis and highlight the crucial role of m6A modification and its modulators in adipogenesis.

Swine enteric coronaviruses (SECoVs), including porcine epidemic diarrhea virus (PEDV), swine acute diarrhea syndrome coronavirus (SADS-CoV), and porcine deltacoronavirus (PDCoV) have emerged and been prevalent in pig populations in China for the last several years. However, current traditional inactivated and attenuated PEDV vaccines are of limited efficacy against circulating PEDV variants, and there are no commercial vaccines for prevention of PDCoV and SADS-CoV. RNA interference (RNAi) is a powerful tool in therapeutic applications to inhibit viral replication in vitro. In this study, we developed a small interfering RNA generation system that expressed two different short hairpin RNAs (shRNAs) targeting the M gene of PEDV and SADS-CoV and the N gene of PDCoV, respectively. Our results demonstrated that simultaneous expression of these specific shRNA molecules inhibited expression of PEDV M gene, SADS-CoV M gene, and PDCoV N gene RNA by 99.7%, 99.4%, and 98.8%, respectively, in infected cell cultures. In addition, shRNA molecules significantly restricted the expression of M and N protein, and impaired the replication of PEDV, SADS-CoV, and PDCoV simultaneously. Taken together, this shRNAs expression system not only is proved to be a novel approach for studying functions of various genes synchronously, but also developed to test aspects of a potential therapeutic option for treatment and prevention of multiple SECoV infections., Highlights • Two potential targets of antiviral molecules for the treatment of three swine enteric coronaviruses replication are tested. • The multiple-shRNA expression vector was constructed to effectively inhibit PEDV, SADS-CoV, and PDCoV. • A shRNA-based multiple-resistance antiviral strategy, significantly affecting viral replication, was evaluated in vitro.

Bone marrow stem cells (BMSCs) are multipotent stem cells that can regenerate mesenchymal tissues, such as adipose tissue, bone, and muscle. Recent studies have shown that N6-methyladenosine (m6A) methylation, one of the most prevalent epigenetic modifications, is involved in the development process. However, whether it plays roles in BMSC differentiation is still elusive. Here, we found that the deletion of m6A "writer" protein methyltransferase-like (METTL)3 in porcine BMSCs (pBMSCs) could promote adipogenesis and janus kinase (JAK)1 protein expression via an m6A-dependent way. Knockdown of METTL3 decreased mRNA m6A levels of JAK1, leading to enhanced YTH m6A RNA binding protein 2 (YTHDF2)-dependent JAK1 mRNA stability. We further demonstrated that JAK1 activated signal transducer and activator of transcription (STAT) 5 through regulation of its phosphorylation to bind to the promoter of CCAAT/enhancer binding protein (C/EBP) β, which could ultimately lead to a modulated adipogenic process. Collectively, our results reveal an orchestrated network linking the m6A methylation and JAK1/STAT5/C/EBPβ pathway in pBMSCs adipogenic differentiation. Our findings provide novel insights into the underlying molecular mechanisms of m6A modification in the regulation of BMSCs differentiating into adipocytes, which may pave a way to develop more effective therapeutic strategies in stem cell regenerative medicine and the treatment of obesity.-Yao, Y., Bi, Z., Wu, R., Zhao, Y., Liu, Y., Liu, Q., Wang, Y., Wang, X. METTL3 inhibits BMSC adipogenic differentiation by targeting the JAK1/STAT5/C/EBPβ pathway via an m6A-YTHDF2-dependent manner.

N6-methyladenosine (m6A) is the most abundant-internal modification of eukaryotic mRNA. m6A can be installed and removed by specific enzymes. The "writer," "eraser," and "reader" of m6A modification have been reported. These discoveries facilitate our understanding of the functional significance of m6A. m6A plays an essential role in diverse biological processes by recruiting the corresponding YTH domain-containing proteins, as well as recruiting additional translation initiation factors. Here, we provide an update on the various aspects of YTH domain-containing proteins, including an introduction to the YTH domain, the categories, distribution in cells, and biological roles of YTH proteins. Then we focus on the mechanisms that YTH proteins recognize m6A and mediate the fate of methylated-RNAs in eukaryotic cells.

An efficient and stable combined denitrification-partial nitrification-Anammox process with a dual recycling system was used to remove nitrogen from mature landfill leachate. After 155 d of operation, the NO3− as the PN-Anammox byproduct was almost treated with biodegradable organic carbon in raw wastewater in a pre-denitrification reactor by external recycling system. When raw landfill leachate with NH4+-N concentration of 1900 mg/L was treated, an integrated reactor with airlift recycling was combined with the PN and Anammox reactions to efficiently remove NH4+ from the inflow. The total nitrogen concentration of effluent stabilized at 20 mg/L and total nitrogen removal efficiency was 99%. The maximum NO2− production rate in the aerobic zone was 2.2 kg/(m3·d) and the maximum nitrogen removal rate in the anaerobic zone was 21.4 kg/(m3·d). The most common phyla among the nitrification and the Anammox functional bacteria were Nitrosomonas, Candidatus Kuenenia, and Candidatus Brocadia after landfill leachate treatment.

N6-methyladenosine (m6A) is the most prevalent internal mRNA modification in eukaryotes. Loss of m6A demethylase FTO increases m6A levels and inhibits adipogenesis of preadipocytes. However, its underlying mechanism remains elusive. Here, we demonstrated that silencing FTO inhibited adipogenesis of preadipocytes through impairing cell cycle progression at the early stage of adipogenesis. FTO knockdown markedly decreased the expression of CCNA2 and CDK2, crucial cell cycle regulators, leading to delayed entry of MDI-induced cells into G2 phase. Furthermore, the m6A levels of CCNA2 and CDK2 mRNA were significantly upregulated following FTO knockdown. m6A-binding protein YTHDF2 recognized and decayed methylated mRNAs of CCNA2 and CDK2, leading to decreased protein expression, thereby prolonging cell cycle progression and suppressing adipogenesis. Our work unravels that FTO regulates adipogenesis by controlling cell cycle progression in an m6A-YTHDF2 dependent manner, which provides insights into critical roles of m6A methylation in adipogenesis.

The phosphorus recovery efficiency in PAOs-biofilm system is so far limited to stimulating phosphorus release by adding concentrated organic carbon solution during the anaerobic stage. In present study, a PAOs-biofilm sequence batch reactor (BSBR) were operated to investigate whether increase of the phosphorus storage content via DO control can stimulate the release in the biofilm. During the operation of BSBR for 160 days, the phosphorus content in biofilm (Pbiofilm) was doubled via increasing dissolved oxygen (DO) from 4 to 6 mg/L. With the COD of 200 mg/L in the anaerobic phase, the phosphorus release was enhaced, resulting in an significant increase of phosphorus concentration from 94.85 to 179.5 mg/L in recirculated solution. Batch tests further clarified explicitly the increase of Pbiofilm stimulated a phosphorus release rate but this must be balanced since high Pbiofilm reduced the phosphorus removal capacity of the biofilm. With analysis of P31-NMR, Ortho-P and Poly-P were the main phosphorus species stored in biofilm. The microbial cell played a more important role than EPS in phosphorus storage. The dominant phylum in the master reactor was Proteobacteria with an abundance of 64.4%, in which the Rhodocyclaceae was the dominant PAOs with an abundance of 10.1%. The outcome of this study elucidated that increase of phosphorus storage via DO control which facilitated more efficient phosphorus release with much lower organic carbon source consumption during the enrichment of phosphorus. Meanwhile, we provided a new perspective for the recovery of phosphorus in future wastewater treatment plants.

The effects of salinity changes on nitrogen transformation efficiency and recoverability were studied by using a partial nitration (PN)- anaerobic ammonium oxidation (Anammox) integrated reactor. The changes of microbial community structure and population abundance during the increase and decrease of salinity were also analyzed by 16S rRNA gene high-throughput sequencing. The results showed that when the salinity was increased to 1.35%, the combined PN-Anammox process achieved the maximum stimulated and total nitrogen removal rate (TNRR) arrived at 1.1kg/(m3·d). When the salinity was higher than 1.35%, the activities of AOB and Anammox bacteria began to be inhibited. When the salinity reached 2.4%, the TNRR decreased to 60%. TNRR was fast restored, when salinity was reduced to 0.11%. The genes of AOB and Anammox bacteria indicated that the TNRR of the reactor was restored after salinity inhibition, but the functional microbial community structure and abundance had relatively large, irreversible changes.

In this work, a novel nitrate (NO3−) reduction pathway by anaerobic ammonium oxidation (anammox) biomass was firstly discovered with the intracellular carbon sources as the only electron donors. And the possible reaction mechanism was deduced to be intracellular dissimilatory nitrate reduction to ammonium (DNRA) pathway according to the experimental results. In batch experiments, without any external electron donors, NO3−-N (about 50 mg/L) was reduced to N2 within 48 h, and a small amount of NO2−-N was detected (the maximum of 2 mg/L) with the anammox biomass concentration of 4400 mg/L. Acetylene (4.46 mmol/L) addition resulted in obvious NH4+ accumulation during NO3− degradation by anammox biomass, since acetylene mainly interfered in hydrazine (N2H4) generation from NH4+ and NO. Without HCO3− addition, the NO3−-N degradation rate was slower than that with HCO3− addition. Simultaneously, glycogen contents inside anammox biomass decreased to 133.22 ± 1.21 mg/g VSS and 129.79 ± 1.21 mg/g VSS with and without HCO3−, respectively, from 142.20 ± 0.61 mg/g VSS. In the long-term experiment, anammox biomass stably degraded NO3−-N without external electron donors addition, and the maximum removal efficiency of NO3−-N reached 55.4%. The above results indicated the anammox bacteria utilized the DNRA pathway to reduce NO3− to NO2− and further NH4+, then normal anammox metabolism would continue to convert the produced NO2− and NH4+ to N2. The intracellular stored carbon sources (e.g., glycogen) were supposed to be electron donors for NO3− degradation. This capability would enhance the viability and living space of anammox bacteria in different natural ecosystems, and make it plausible that complete nitrogen removal could be implemented only by the anammox process.

Simple Summary Porcine deltacoronavirus (PDCoV) is an emerging enteropathogen distributed globally, which causes substantial economic losses in the swine industry. The characterization of the receptor promiscuity may pose a risk of cross-species transmission. However, the options for pharmaceutical interventions are limited. In this study, the vectors expressing shRNAs targeting the nucleocapsid gene were generated to assess the inhibition effect of PDCoV reproduction. Our preliminary results demonstrate that a dual shRNA expression system is an effective strategy in combating PDCoV infection without cytotoxicity, which would facilitate the ongoing development of RNAi-based therapeutic drugs against viral diseases. Abstract Porcine deltacoronavirus (PDCoV) is a novel enteric coronavirus and is becoming one of the major causative agents of diarrhea in pig herds in recent years. To date, there are no commercial vaccines or antiviral pharmaceutical agents available to control PDCoV infection. Therefore, developing a reliable strategy against PDCoV is urgently needed. In this study, to observe the antiviral activity of RNA interference (RNAi), four short hairpin RNAs (shRNAs) specific to the nucleocapsid (N) gene of PDCoV were designed and tested in vitro. Of these, a double-shRNA-expression vector, designated as pSil-double-shRNA-N1, was the most effectively expressed, and the inhibition of PDCoV replication was then further evaluated in neonatal piglets. Our preliminary results reveal that plasmid-based double-shRNA-expression targeting the N gene of PDCoV can significantly protect LLC-PK1 cells and piglets from pathological lesions induced by PDCoV. Our study could benefit the investigation of the specific functions of viral genes related to PDCoV infection and offer a possible methodology of RNAi-based therapeutics for PDCoV infection.

For municipal wastewater with low temperature and ammonium, conventional oxygen-limited have difficulty achieving long-term stable inhibition of nitrite oxidizing bacteria (NOB) and stable nitritation. So a partial nitrification-anaerobic ammonium oxidation integrated reactor with independent partitions was used to investigate the feasibility of adding an auto-recycling system to promote low exposure of nitrite in the aerobic zone and to inhibit the NOB activity. The results showed that nitrite produced in the aerobic zone could be timely transported to the anaerobic zone for Anammox utilization, and the nitrite nitrogen concentration was diluted to keep within 1 mg/L in the aerobic zone by the effluent recycling. NOB growth was inhibited by nitrite deficiency. The maximum nitrogen removal rate of the reactor was 0.29 kg/(m3·d), and the nitrate nitrogen production rate of NOB was controlled within 0.04 kg/(m3·d). Nitrosomonas and Candidatus Kuenenia were found as functional species of ammonia-oxidizing bacteria and Anammox bacteria, respectively.

Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) hold great promise for regenerative medicine, disease treatment, and organ transplantation. As the ethical issue of human ESCs and similarity of pig in human genome and physiological characteristics, the porcine iPSCs (piPSCs) have become an ideal alternative study model. N6-methyladenosine (m6A) methylation is the most prevalent modification in eukaryotic mRNAs, regulating the self-renewal and differentiation of pluripotency stem cells. However, the explicit m6A-regulating machinery remains controversial. Here, we demonstrate that m6A modification and its modulators play a crucial role in mediating piPSCs pluripotency. In brief, loss of METTL3 significantly impairs self-renewal and triggers differentiation of piPSCs by interfering JAK2 and SOCS3 expression, further inactivating JAK2–STAT3 pathway, which then blocks the transcription of KLF4 and SOX2. We identify that both of JAK2 and SOSC3 have m6A modification at 3′UTR by m6A-seq analysis. Dual-luciferase assay shows that METTL3 regulates JAK2 and SOCS3 expression in an m6A-dependent way. RIP-qPCR validates JAK2 and SOCS3 are the targets of YTHDF1 and YTHDF2, respectively. SiMETTL3 induced lower m6A levels of JAK2 and SOCS3 lead to the inhibition of YTHDF1-mediated JAK2 translation and the block of YTHDF2-dependent SOCS3 mRNA decay. Subsequently, the altered protein expressions of JAK2 and SOCS3 inhibit JAK2–STAT3 pathway and then the pluripotency of piPSCs. Collectively, our work uncovers the critical role of m6A modification and its modulators in regulating piPSCs pluripotency and provides insight into an orchestrated network linking the m6A methylation and SOCS3/JAK2/STAT3 pathway in pluripotency regulation.

Highlights • RT-LAMP assay was established for the simple and rapid detection of PPgV. • RT-LAMP assay showed strong specificity with high sensitivity. • RT-LAMP assay provides an extremely valuable screening tool for the surveillance of PPgV in field conditions., A simple and accurate reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was developed and evaluated for the detection of porcine pegivirus (PPgV). The specific RT-LAMP primers targeting the conserved regions of NS5A genes were designed and used to detect PPgV. The optimal reaction parameter for RT-LAMP assay was 63℃ for 60 min. The detection limit of the RT-LAMP assay was 10 copies of PPgV genome, which was 100 times more sensitive than that of the conventional RT-PCR and comparable to nested RT-PCR and quantitative RT-PCR (qRT-PCR). There was no cross amplification with other related RNA viruses. In the clinical evaluation, the RT-LAMP assay exhibited a similar sensitivity with nested RT-PCR and qRT-PCR. The results indicated that RT-LAMP assay developed in this study could be a highly specific, sensitive, and cost-effective alternative for a rapid detection of PPgV in field settings.

The full-length genome sequence of a novel swine acute diarrhea syndrome coronavirus (SADS-CoV), CH/FJWT/2018, was determined, which was genetically most closely related to CN/GDWT/2017, recently discovered in Fujian, China. The indel sites of the spike (S) gene of CH/FJWT/2018 were most similar to those of bat-origin SADS-related coronaviruses.

The differences in the physiological characteristics between AOB and ANAMMOX bacteria lead to suboptimal performance when used in a single reactor. In this study, aerobic and anaerobic zones with different survival environments were constructed in a single reactor to realize partitioned culture of AOB and ANAMMOX bacteria. An external air-lift reflux system was formed which used the exhaust from the aeration zone as power to return the effluent to the aeration zone. The reflux system effectively alleviated the large pH fluctuations and promoted NO2--N to rapidly use by ANAMMOX bacteria, effectively inhibiting the activity of NOB. After 95d of running, the nitrogen removal rate increased from the initial 0.21kg/(m3·d) to 3.1kg/(m3·d). FISH analyses further demonstrated that AOB and ANAMMOX bacteria acquired efficient enrichment in the corresponding zone. Thus, this type of integrative reactor may create the environments needed for the partial nitritation-ANAMMOX processing.

This study investigated the inhibitory effects of Cd, Ag, Hg and Pb on Anammox biomass by batch experiments. Inhibition effects and recovery performance were monitored in sequence over 24 h and 72 h experimental runs. The IC50 (the 50% inhibitory concentration of nitrogen removal rate) values were calculated as follows: 11.16 ± 0.42 mg/L for Cd; 11.52 ± 0.49 mg/L for Ag; 60.35 ± 2.47 mg/L for Hg. Pb caused only 7.19% decrease on nitrogen removal rate (NRR) even at a dose of 40 mg/L. Hence, the order of the inhibitory effect was determined as Cd > Ag > Hg > Pb. Furthermore, the crude enzyme (hydrazine dehydrogenase) activity and heme c concentration were also depressed under heavy metals exposure. Among four metals, Cd and Hg could exert sustained toxicity on Anammox biomass, but mal-effects of Ag and Pb could resume to a large extent after removing these two metals within 96 h.

Agrobacterium vitis strain E26 is a promising biocontrol agent of grapevine crown gall, an economically important disease of grape worldwide. In this report, we developed a Plating-PCR method that allows specific detection and quantification of E26 by combining classical microbiological techniques with molecular tools. Random amplified polymorphic DNA fingerprints were used to differentiate E26 from other A. vitis strains. A differentially amplified fragment from E26 was sequenced and characterized as a sequence characterized amplified region (SCAR) marker. Two primer pairs were then designed and evaluated for their specificity against E26. One of the two SCAR primer pairs, 740F/R, was further selected for specific detection of strain E26. A plating assay coupled to PCR with the SCAR primers 740F/R allowed the assessment of population dynamics of E26 in non-sterile grape rhizosphere soil under controlled conditions.

Spotted Halibut (Verasper variegatus) is a commercially important marine fish species. In the present study, to isolate sex-specific markers in Spotted Halibut, we screened the genomes of Spotted Halibut by AFLP technique with 64 different primer combinations. Two primer combinations, MseI-CAG/EcoRI-ACC and MseI-CAT/EcoRI-AGG, produced a female-specific fragment in all females (n = 88) and in no males (n = 60, except 3 individuals), respectively. Both fragments were excised from the gel, cloned and characterized. The first fragment (named VevaF533) was 533 bp long, while the length of the second one (named VevaF218) was 218 bp. The two sequences showed no similarity to each other, and to the known sequences existing in the GenBank database using BLASTn. Cross-species amplification showed that the marker VevaF218 is a species-specific marker which is present in Spotted Halibut females but absent in Barfin Flounder (Verasper moseri). So this marker could be used for discriminating unambiguously between Spotted Halibut females and Barfin Flounder. Examination of the patterns of inheritance of VevaF218 in an interspecific hybrid family (V. variegatus ♂×V. moseri ♀) showed a female-specific pattern of inheritance from mother to daughter, implying that the marker VevaF218 is located on the female sex chromosome. This study provides a reliable AFLP-based genetic sexing of Spotted Halibut that could be useful for genetic mapping of the sex chromosomes and identification of sex-linked genes.

Two female-specific AFLP (amplified fragment length polymorphism) markers (named CseF464 and CseF136) were isolated by using one selective primer combination (E-AGC/M-CTG) from the genomic DNA of 20 females and 20 males of the half-smooth tongue sole Cynoglossus semilaevis. Both the markers were re-amplified, recovered from the agarose gels, cloned and sequenced. Bioinformatics analysis indicated that the length of the two markers were 468 bp and 134 bp, respectively, and the sequences showed no similarity to each other, as well as to the known sequences deposited in the GenBank database using BLASTn. Two pairs of SCAR (sequence characterized amplified regions) primers were designed based on the sequences of the two female-specific markers. Furthermore, PCR-based genetic sex identification method was developed in Cynoglossus semilaevis. A specific fragment was amplified in all females but not in any males by using these SCAR primers on the initial 20 female and 20 male individuals of Cynoglossus semilaevis. The feasibility of the two SCAR primer pairs was confirmed in additional 100 individuals (50 females and 50 males). This allowed for reliable, rapid molecular identification of genetic sex of the species, genetic mapping on the sex chromosomes and better understanding of the sex determination and sex differentiation in the half smooth tongue sole.

Ribose methylation of eukaryotic rRNA is directed by box C/D small nucleolar RNAs (snoRNAs), which pinpoint the nucleotide to be methylated in specific position within the rRNA sequence. Here, we report the identification of a novel double-guide C/D box snoRNA termed snR88 that directs methylation of two previously undetermined sites in 25S rRNA from the fission yeast. Knockout of the predicted TATA box of the snR88 gene resulted in the complete blocking of its expression, showing that snR88 is an independently transcribed gene and dispensable for yeast viability. The depletion of snR88 abolished 25S rRNA methylation at U2304 and U2497 simultaneously. Interestingly, an unusual pause of reverse transcription at U2495 was observed, which implies an unknown structure of 25S rRNA related to ribose methylation at U2497 in the fission yeast.

Plasma membrane intrinsic proteins (PIPs) are one subfamily of aquaporins that mediate the transmembrane transport of water. To reveal their function in poplar, we generated transgenic poplar plants in which the translation of PIP genes was downregulated by RNA interference investigated these plants with a comprehensive leaf plasma membrane proteome and physiome analysis. First, inhibition of PIP synthesis strongly altered the leaf plasma membrane protein composition. Strikingly, several signaling components and transporters involved in the regulation of stomatal movement were differentially regulated in transgenic poplars. Furthermore, hormonal crosstalk related to abscisic acid, auxin and brassinosteroids was altered, in addition to cell wall biosynthesis/cutinization, the organization of cellular structures and membrane trafficking. A physiological analysis confirmed the proteomic results. The leaves had wider opened stomata and higher net CO2 assimilation and transpiration rates as well as greater mesophyll conductance for CO2 (gm) and leaf hydraulic conductance (Kleaf). Based on these results, we conclude that PIP proteins not only play essential roles in whole leaf water and CO2 flux but have important roles in the regulation of stomatal movement. Biological significance The present work is a comprehensive survey combining leaf plasma membrane proteomics and physiological methods to assess the functionality of the whole PIP subfamily in tree model system.

Transcriptome studies have revealed that many non-coding RNAs (ncRNAs) are located near the 3' sense terminus of protein-coding genes. However, the transcription and function of these RNAs remain elusive. Here, we identify a 3' sense termini-associated sRNA (TASR) downstream of rpl26 in Schizosaccharomyces pombe (S. pombe). Structure and function assays indicate that the TASR is an H/ACA box snoRNA required for 18S rRNA pseudouridylation at U121 and U305 sites and is therefore a cognate of snR49 from the budding yeast. Transcriptional studies show that pre-snR49 overlaps most of the coding sequence (CDS) of rpl26. Using scanning deletion analysis within promoter region, we show that the rpl26 promoter is required for the 3' TASR transcription. Interestingly, chromosomal synteny of rpl26-snR49 is found in the Schizosaccharomyces groups. Taken together, we have revealed a new transcriptional mechanism for 3' sense TASRs, which are transcribed by the same promoter as their upstream protein genes. These results further suggest that the origin and function of 3' sense ncRNAs are associated with upstream genes in higher eukaryotes.

The cutlassfish (Trichiurus haumela) is an important commercial fish species. In the present study, we report 12 polymorphic microsatellite DNA markers for cutlassfish. The number of alleles per locus ranged from 2 to 9 in a sample of 26 individuals. Observed and expected heterozygosities per locus varied from 0.2727 to 0.9583 and from 0.4059 to 0.7926, respectively. Two loci (Trha25 and Trha27) showed significant departure from Hardy–Weinberg equilibrium after sequential Bonferroni correction (P < 0.0042). No significant linkage disequilibrium between pairs of loci was found. These microsatellite markers provide powerful tools for investigating genetic population structure, population history and conservation management of cutlassfish.

In this study, we explored the effects of Fe(II) on the activity of anammox biomass both in short and long terms. With the influent Fe(II) at 0.09 and 0.12 mM, continuous experiments demonstrated that the nitrogen removal rates increased 32.2% and 29.9% compared to that with normal Fe(II) level (0.03 mM). Influent Fe(II) variation could affect the total Fe, heme c levels and hydrazine dehydrogenase activity. At the Fe(II) concentration of 0.09 mM, the total Fe, heme c levels inside anammox cell and hydrazine dehydrogenase activity could increase about 2.0, 2.1 and 2.35 folds as much as that with normal Fe(II) level. However, when the Fe(II) elevated to 0.18 mM, it would cause a mal-effect on anammox bacteria and further deteriorate nitrogen removal performance. It was indicated that the appropriate increase of Fe(II) was beneficial for more heme c synthesis, enhancement of hydrazine dehydrogenase activity, the growth of anammox bacteria.

This study examined long-term effect of MnO(2) powder (average diameter of 4-7 μm) on nitrogen removal in anammox process. Two lab-scale up-flow anammox reactors were operated for 380 days, one with and one without MnO(2) powder addition. During the period when only substrate concentrations varied, the maximum nitrogen removal rate in the reactor with MnO(2) addition reached 920.9 g-N/m(3)/d. This value was 2-folds higher than that (464.6 g-N/m(3)/d) of the reactor without MnO(2) addition. The crude enzyme activities of the anammox biomass from the two reactors was measured as 0.531±0.019 and 0.298±0.007 μmol cytochrome c reduced/mg protein/min, respectively. Transmission electron microscopy observation demonstrated more undefined particles existing inside anammox bacterial cell in the reactor with MnO(2) powder addition. Furthermore, filament-like structures inside anammoxosome were observed, which formed a net-like structure with particles as the connecting nodes. The experiment results demonstrated that MnO(2) improved nitrogen removal performance of anammox process.

indicus). Conversely, the allele frequency of D12 was extremely low compared with other breeds (ranging from 0.14 in German Brown to 0.74 in Japanese Holstein). Moreover, we observed four haplotypes in Chinese yak with respect to both indel loci, and the haplotype frequency of D23–D12 was very low. Interestingly, although the frequency distributions of allele, genotype and haplotype showed no statistical difference among the surveyed populations, the distribution patterns in Chinese yak were significantly different from those in BSE-affected cattle (P < 0.03 in each comparison).