Allocation patterns of nonstructural carbohydrates in response to CO_(2)elevation and nitrogen deposition in Cunninghamia lanceolata saplings

Stored nonstructural carbohydrates(NSC)indicate a balance between photosynthetic carbon(C)assimilation and growth investment or loss through respiration and root exudation.They play an important role in plant function and whole-plant level C cycling.CO_(2)elevation and nitrogen(N)deposition,which are two major environmental issues worldwide,aff ect plant photosynthetic C assimilation and C release in forest ecosystems.However,information regarding the eff ect of CO_(2)elevation and N deposition on NSC storage in diff erent organs remains limited,especially regarding the trade-off between growth and NSC reserves.Therefore,here we analyzed the variations in the NSC storage in diff erent organs of Chinese fi r(Cunninghamia lanceolata)under CO_(2)elevation and N addition and found that NSC concentrations and contents in all organs of Chinese fi r saplings increased remarkably under CO_(2)elevation.However,N addition induced diff erential accumulation of NSC among various organs.Specifi cally,N addition decreased the NSC concentrations of needles,branches,stems,and fi ne roots,but increased the NSC contents of branches and coarse roots.The increase in the NSC contents of roots was more pronounced than that in the NSC content of aboveground organs under CO_(2)elevation.The role of N addition in the increase in the structural biomass of aboveground organs was greater than that in the increase in the structural biomass of roots.This result indicated that a diff erent tradeoff between growth and NSC storage occurred to alleviate resource limitations under CO_(2)elevation and N addition and highlights the importance of separating biomass into structural biomass and NSC reserves when investigating the eff ects of environmental change on biomass allocation.

Polyphosphate Accelerates Transformation of Nonstructural Carbohydrates to Improve Growth of ppk-Expressing Transgenic Rice in Phosphorus Deficiency Culture

Crop yield and quality are often limited by the amount of phosphate fertilizer added to infertile soils,a key limiting factor for sustainable development in modern agriculture.The polyphosphate kinase(ppk)gene-expressing transgenic rice with a single-copy line(ETRS)is constructed to improve phosphate fertilizer utilization efficiency for phosphorus resource conservation.To investigate the potential mechanisms of the increased biomass in ETRS in low phosphate culture,ETRS was cultivated in a low inorganic phosphate(Pi)culture medium(15μmol/L Pi,LP)and a normal Pi culture medium(300μmol/L Pi,CP),respectively.After 89 d of cultivation in different concentrations of phosphate culture media,the total phosphorus,polyphosphate(polyP),biomass,photosynthetic rate,nonstructural carbohydrate(NSC)contents,related enzyme activities,and related gene expression levels were analyzed.The results showed that ETRS had a high polyP amount to promote the photosynthetic rate in LP,and its biomass was almost the same as the wild type(WT)in CP.The NSC content of ETRS in LP was higher than that of WT in LP,but slightly lower than that of WT in CP.PolyP notably promoted the sucrose phosphate synthase activities of ETRS and significantly down-regulated the expression levels of sucrose transporter genes(OsSUT3 and OsSUT4),resulting in inhibiting the transport of sucrose from shoot to root in ETRS.It was concluded that polyP can stimulate the synthesis of NSCs in LP,which improved the growth of ETRS and triggered the biological activities of ETRS to save phosphate fertilizer.Our study provides a new way to improve the utilization rate of phosphate fertilizer in rice production.

Calcitriol attenuates liver fibrosis through hepatitis C virus nonstructural protein 3-transactivated protein 1-mediated TGF β1/Smad3 and NF-κB signaling pathways

BACKGROUND Hepatic fibrosis is a serious condition,and the development of hepatic fibrosis can lead to a series of complications.However,the pathogenesis of hepatic fibrosis remains unclear,and effective therapy options are still lacking.Our group identified hepatitis C virus nonstructural protein 3-transactivated protein 1(NS3TP1) by suppressive subtractive hybridization and bioinformatics analysis,but its role in diseases including hepatic fibrosis remains undefined.Therefore,additional studies on the function of NS3TP1 in hepatic fibrosis are urgently needed to provide new targets for treatment.AIM To elucidate the mechanism of NS3TP1 in hepatic fibrosis and the regulatory effects of calcitriol on NS3TP1.METHODS Twenty-four male C57BL/6 mice were randomized and separated into three groups,comprising the normal,fibrosis,and calcitriol treatment groups,and liver fibrosis was modeled by carbon tetrachloride(CCl4).To evaluate the level of hepatic fibrosis in every group,serological and pathological examinations of the liver were conducted.TGF-β1 was administered to boost the in vitro cultivation of LX-2 cells.NS3TP1,α-smooth muscle actin(α-SMA),collagen I,and collagen Ⅲ in every group were examined using a Western blot and real-time quantitative polymerase chain reaction.The activity of the transforming growth factor beta 1(TGFβ1)/Smad3 and NF-κB signaling pathways in each group of cells transfected with pcDNA-NS3TP1 or siRNA-NS3TP1 was detected.The statistical analysis of the data was performed using the Student’s t test.RESULTS NS3TP1 promoted the activation,proliferation,and differentiation of hepatic stellate cells(HSCs)and enhanced hepatic fibrosis via the TGFβ1/Smad3 and NF-κB signaling pathways,as evidenced by the presence of α-SMA,collagen I,collagen Ⅲ,p-smad3,and p-p65 in LX-2 cells,which were upregulated after NS3TP1 overexpression and downregulated after NS3TP1 interference.The proliferation of HSCs was lowered after NS3TP1 interference and elevated after NS3TP1 overexpression,as shown by the luciferase assay.NS3TP1 inhibited the apoptosis of HSCs.Moreover,both Smad3 and p65 could bind to NS3TP1,and p65 increased the promoter activity of NS3TP1,while NS3TP1 increased the promoter activity of TGFβ1 receptor I,as indicated by coimmunoprecipitation and luciferase assay results.Both in vivo and in vitro,treatment with calcitriol dramatically reduced the expression of NS3TP1.Calcitriol therapy-controlled HSCs activation,proliferation,and differentiation and substantially suppressed CCl4-induced hepatic fibrosis in mice.Furthermore,calcitriol modulated the activities of the above signaling pathways via downregulation of NS3TP1.CONCLUSION Our results suggest that calcitriol may be employed as an adjuvant therapy for hepatic fibrosis and that NS3TP1 is a unique,prospective therapeutic target in hepatic fibrosis.

Interannual dynamics of stemwood nonstructural carbohydrates in temperate forest trees surrounding drought

Interactions between water and carbon dynamics underlie drought-related tree mortality.While whole-tree water relations have been shown to play a key role in the response to and recovery from drought,the role of nonstructural carbohydrates(NSC) and how their storage and allocation changes surrounding drought events deserves further attention and is critical for understanding tree survival.Here,we quantified in situ NSC responses of temperate forest trees to the 2016 drought in the northeastern United States.Sugar and starch concentrations were measured in the stemwood of five tree species from 2014 to 2019,which allowed us to monitor NSCs in relation to climatic conditions before,during,and after the natural drought.We found that immediately following the drought,measured stemwood NSC concentrations decreased.However,NSC concentrations rebounded quickly within three years.Notably,trees allocated proportionally more to starch than to sugars following the 2016 drought.In winter 2017,starch comprised 45% of total stemwood stores,whereas starch made up only 1-2% in other years.Further,we modeled and assessed the climatic drivers of total NSC concentrations in the stem.Variation in total NSC concentrations was significantly predicted by the previous year’s temperature,precipitation,and standardized precipitation-evapotranspiration index(SPEI),with stemwood concentrations decreasing following hotter,drier periods and increasing following cooler,wetter periods.Overall,our work provides insight into the climatic drivers of NSC storage and highlights the important role that a tree’s carbon economy may play in its response and recovery to environmental stress.

Performance-based seismic design of nonstructural building components:The next frontier of earthquake engineering

With the development and implementation of performance-based earthquake engineering,harmonization of performance levels between structural and nonstructural components becomes vital. Even if the structural components of a building achieve a continuous or immediate occupancy performance level after a seismic event,failure of architectural,mechanical or electrical components can lower the performance level of the entire building system. This reduction in performance caused by the vulnerability of nonstructural components has been observed during recent earthquakes worldwide. Moreover,nonstructural damage has limited the functionality of critical facilities,such as hospitals,following major seismic events. The investment in nonstructural components and building contents is far greater than that of structural components and framing. Therefore,it is not surprising that in many past earthquakes,losses from damage to nonstructural components have exceeded losses from structural damage. Furthermore,the failure of nonstructural components can become a safety hazard or can hamper the safe movement of occupants evacuating buildings,or of rescue workers entering buildings. In comparison to structural components and systems,there is relatively limited information on the seismic design of nonstructural components. Basic research work in this area has been sparse,and the available codes and guidelines are usually,for the most part,based on past experiences,engineering judgment and intuition,rather than on objective experimental and analytical results. Often,design engineers are forced to start almost from square one after each earthquake event: to observe what went wrong and to try to prevent repetitions. This is a consequence of the empirical nature of current seismic regulations and guidelines for nonstructural components. This review paper summarizes current knowledge on the seismic design and analysis of nonstructural building components,identifying major knowledge gaps that will need to be filled by future research. Furthermore,considering recent trends in earthquake engineering,the paper explores how performance-based seismic design might be conceived for nonstructural components,drawing on recent developments made in the field of seismic design and hinting at the specific considerations required for nonstructural components.

Quantifying the effects of short-term heat stress at booting stage on nonstructural carbohydrates remobilization in rice

Extreme heat stress events are becoming more frequent under anticipated climate change,which can have devastating impacts on rice growth and yield.To quantify the effects of short-term heat stress at booting stage on nonstructural carbohydrates(NSC)remobilization in rice,two varieties(Nanjing 41 and Wuyunjing 24)were subjected to 32/22/27°C(maximum/minimum/mean),36/26/31°C,40/30/35°C,and 44/34/39°C for 2,4 and 6 days in phytotrons at booting stage during 2014 and 2015.Yield and yield components,dry matter partitioning index(DMPI),NSC accumulation and translocation were measured and calculated.The results showed that the increase of high-temperature level and duration significantly reduced grain yield by suppressing spikelet number per panicle,seed-setting rate,and grain weight.Heat stress at booting decreased DMPI in panicles,increased DMPI in stems,but had no significant effect on photosynthetic rate.Stem NSC concentration increased whereas panicles NSC concentration,stem NSC translocation efficiency,and contribution of stem NSC to grain yield decreased.Severe heat stress even transformed the stem into a carbohydrate sink during grain filling.The heat-tolerant Wuyunjing 24 showed a higher NSC transport capacity under heat stress than the heat-sensitive Nanjing 41.Heat degree-days(HDD),which combines the effects of the intensity and duration of heat stress,used for quantifying the impacts of heat stress indicates the threshold HDD for the termination of NSC translocation is 9.82°C day.Grain yield was negatively correlated with stem NSC concentration and accumulation at maturity,and yield reduction was tightly related to NSC translocation reduction.The results suggest that heat stress at booting inhibits NSC translocation due to sink size reduction.Therefore,genotypes with higher NSC transport capacity under heat stress could be beneficial for rice yield formation.

Function of nonstructural 5A protein of genotype 2a in replication and infection of HCV with gene substitution

AIM:To explore the function of Nonstructural 5A(NS5A) protein of genotype 2a(JFH1)in the replication and infection of hepatitis C virus(HCV).METHODS:Intergenotypic chimera FL-J6JFH/J4NS5A was constructed by inserting NS5A gene from 1b stain HC-J4 by the overlapping polymerase chain reaction (PCR)method and the restriction enzyme reaction.In vitro RNA transcripts of chimera,prototype J6JFH and negative control J6JFH1(GND)were prepared and transfected into Huh-7.5 cells with liposomes.Immunofluorescence assay(IFA),fluorescence quantitative PCR and infection assay were performed to determine the protein expression and gene replication in Huh-7.5 cells.RESULTS:The HCV RNA levels in FL-J6JFH/J4NS5A chimera RNA transfected cells were significantly lower than the wild type at any indicated time point(2.58 ±5.97×106 vs 4.27±1.72×104,P=0.032).The maximal level of HCV RNA in chimera was 5.6±1.8× 104 GE/μg RNA at day 34 after transfection,while the wild type reached a peak level at day 13 which was 126 folds higher(70.65±14.11×105 vs 0.56±0.90 ×105,P=0.028).HCV proteins could also be detected by IFA in chimera-transfected cells with an obviously low level.Infection assay showed that FL-J6JFH/J4NS5A chimera could produce infectious virus particles,ranging from 10±5 ffu/mL to 78.3±23.6 ffu/mL,while that of FL-J6JFH1 ranged from 5.8±1.5×102 ffu/mL to 2.5±1.4×104 ffu/mL.CONCLUSION:JFH1 NS5A might play an important role in the robust replication of J6JFH1.The establishment of FL-J6JFH/J4NS5A provided a useful platform for studying the function of other proteins of HCV.

Dynamics of nonstructural carbohydrates in seagrass Thalassia hemprichii and its response to shading

A field survey was performed to examine nonstructural carbohydrate （NSC） dynamics in seagrass Thalassia hemprichii at the Xincun Bay in southern China. An indoor experiment to investigate the response of NSC in T. hemprichiito shade was conducted. Belowground tissue of T. hemprichiiwas the dominant site of NSC reserves, and soluble sugar was the primary storage compound. The starch content of belowground tissue was lower in high intertidal areas than in low intertidal areas, indicating that the longer air exposure in high intertidal areas resulted in less NSC synthesis and less accumulation of NSC in T. hemprichii. The lowest level of soluble sugar and its proportion to NSC in belowground tissue were observed near the cage culture area, where the nutrient concentration in water and sediment was the highest; while the highest level of that was observed near the coastal shrimp farm, where salinity was the lowest. Soluble sugar in belowground tis- sue showed the following trend： summer〉spring〉winter〉autumn. This corresponded to seasonal changes in the intensity of light. Leaf sugar accumulated during the autumn-winter period, providing a carbon and energy source for flower bud formation and seed germination. Short-term shading decreased NSC accumu- lation. Collectively, these results suggest that nutrient enrichment, freshwater discharge and exposure to air affect NSC dynamics in T. hemprichii. Light intensity, flower bud formation, and seed germination were all found to induce seasonal variations in NSC in T. hemprichii.

Challenge to overcome: Nonstructural protein 5A-P32 deletion in direct-acting antiviral-based therapy for hepatitis C virus

Interferon(IFN)-based therapy for hepatitis C virus(HCV) infection has recently been replaced by IFNfree direct-acting antiviral(DAA)-based therapy, which has been established as a 1^(st) line therapy with high efficacy and tolerability due to its reasonable safety profile. Resistance-associated substitutions(RASs) have been a weakness of DAA-based therapy. For example, combination therapy with daclatasvir and asunaprevir(DCV/ASV) is less effective for HCV genotype 1-infected patients with Y93H as a nonstructural protein 5A RAS. However, the problem regarding RASs has been gradually overcome with the advent of recently developed DAAs, such as sofosbuvir-based regimens or combination therapy with glecaprevir and pibrentasvir. Despite the high efficiency of DAA-based therapy, some cases fail to achieve viral eradication. P32 deletion, an NS5A RAS, has been gradually noticed in patients with DCV/ASV failure. P32 deletion has been sporadically reported and the prevalence of this RAS has been considered to be low in patients with DCV/ASV failure. Thus, the picture of P32 deletion has not been fully evaluated. Importantly, currently-commercialized DAA-based combination therapy was not likely to be effective for patients with P32 deletion. Exploring and overcoming this RAS is essential for antiviral therapy for chronic hepatitis C.

Molecular Characterization of Nonstructural Protein NS38 of Grass Carp Reovirus

Viral nonstructural proteins in both enveloped and non-enveloped viruses play important roles in viral replication. Protein NS38 of Grass carp reovirus (GCRV), has been deduced to be a non-structural protein, and, consistent with other reoviruses, is considered to cooperate with the NS80 protein in viral particle assembly. To investigate the molecular basis of the role of NS38, a complete protein was expressed in E.coli for the first time. It was found that there is a better expression of NS38 induced with IPTG at 28 ℃ rather than 37 ℃. In addition, the antiserum of NS38 prepared with purified fusion protein and injected into rabbit could be used for detecting NS38 protein expression in GCRV infected cell lysate, while there is not any reaction crossed with purified virus particle, confirming NS38 is not a component of the viral structural protein. The result reported in this study will provide evidence for further viral protein-protein and protein-RNA interaction in dsRNA viruses replication.

Divergent allocations of nonstructural carbohydrates shape growth response to rainfall reduction in two subtropical plantations

Nonstructural carbohydrates(NSC)are indicators of tree carbon balance and play an important role in regulating plant growth and survival.However,our understanding of the mechanism underlying drought-induced response of NSC reserves remains limited.Here,we conducted a long-term throughfall exclusion(TFE)experiment to investigate the seasonal responses of NSC reserves to manipulative drought in two contrasting tree species(a broadleaved tree Castanopsis hystrix Miq.and a coniferous tree Pinus massoniana Lamb.)of the subtropical China.We found that in the dry season,the two tree species differed in their responses of NSC reserves to TFE at either the whole-tree level or by organs,with significantly depleted total NSC reserves in roots in both species.Under the TFE treatment,there were significant increases in the NSC pools of leaves and branches in C.hystrix,which were accompanied by significant decreases in fine root biomass and radial growth without significant changes in canopy photosynthesis;while P.massoniana exhibited significant increase in fine root biomass without significant changes in radial growth.Our results suggested that under prolonged water limitation,NSC usage for growth in C.hystrix is somewhat impaired,such that the TFE treatment resulted in NSC accumulation in aboveground organs(leaf and branch);whereas P.massoniana is capable of efficiently utilizing NSC reserves to maintain its growth under drought conditions.Our findings revealed divergent NSC allocations under experimental drought between the two contrasting tree species,which are important for better understanding the differential impacts of climate change on varying forest trees and plantation types in subtropical China.

Critical role of cytochrome c1 and its cleavage in porcine reproductive and respiratory syndrome virus nonstructural protein 4-induced cell apoptosis via interaction with nsp4

Porcine reproductive and respiratory syndrome virus.（PRRSV） actively induces cell apoptosis both in vitro and in vivo, which can contribute critically to viral pathogenesis. Previous studies have shown that the PRRSV nonstructural protein 4 （nsp4） is an important mediator of this process, but the underlying molecular details remain poorly understood. In this study, we found that the PRRSV nsp4 interacted with the mitochondrial inner membrane protein cytochrome cl （cyto.cl） and induced its proteolytic cleavage. Interestingly, the cleaved N-terminal fragment of cyto.cl was found to exert apoptotic activity, which could cause mitochondrial fragmentation, resulting in apoptotic cell death. And RNA interference （RNAi） silencing experiments further confirmed the crucial role which cyto.cl played in nsp4- and PRRSV-induced cell apoptosis. Thus, our data provide an important piece of mechanistic clues for PRRSV-induced cell apoptosis and also elucidate a novel mechanism for the 3C-like proteases in this finding.

Effects of pruning intensity on nonstructural carbohydrates of Populus alba×P.talassica in the arid desert region of Northwest China

Although pruning is important to obtain highquality,large-diameter timber,the effects of pruning on nonstructural carbohydrates(NSC)in aboveground organs of many timber species are not well understood.Three intensities of pruning(none,moderate and severe)were tested on poplars(Populus alba×P.talassica)in the arid desert region of northwest China to compare the concentrations of soluble sugar(SS),starch(ST)and total nonstructural carbohydrate(TNC)in leaves,branches and trunks during the growing season.The concentration of NSC components after different pruning intensities varied similarly in seasonal patterns,increasing slowly at the beginning of the growing season,continuously declining in the middle,then gradually recovering by the end of the growing season.The monthly mean NSC concentration in poplar differed significantly among the three pruning intensities(p<0.05).The SS concentration in pruned trees was higher than in unpruned trees(p<0.05).For moderately pruned trees,the concentrations of ST and TNC in trunks and branches were higher than in unpruned and in severely pruned trees(p<0.05).Compared with no pruning,pruning changed the seasonal variation in NSC concentration.The orders of SS and TNC concentrations in aboveground organs were leaf>branch>trunk,while the order of ST concentration was trunk>leaf>branch,which was related to functional differences of plant organs.The annual average growth in height of unpruned,moderately pruned,and severely pruned poplars was 0.21±0.06,0.45±0.09 and 0.24±0.05 m,respectively,and the annual average growth in DBH were 0.92±0.04,1.27±0.06 and 1.02±0.05 cm,respectively.Our results demonstrate that moderate pruning may effectively increase the annual growth in tree height and DBH while avoiding damage caused by excessive pruning to the tree body.Therefore,moderate pruning may increase the NSC storage and improve the growth of timber species.

Seismic force demands on acceleration-sensitive nonstructural components:a state-of-the-art review

Nonstructural components(NSCs)are parts,elements,and subsystems that are not part of the primary loadbearing system of building structures but are subject to seismic loading.Damage to NSCs may disrupt the functionality of buildings and result in significant economic losses,injuries,and casualties.In past decades,extensive studies have been conducted on the seismic performance and seismic design methods of NSCs.As the input for the seismic design of NSCs,floor response spectra(FRS)have attracted the attention of researchers worldwide.This paper presents a state-of-the-art review of FRS.Different methods for generating FRS are summarized and compared with those in current seismic design codes.A detailed review of the parameters influencing the FRS is presented.These parameters include the characteristics of ground motion excitation,supporting building and NSCs.The floor acceleration response and the FRS obtained from experimental studies and field observations during earthquakes are also discussed.Three RC frames are used in a case study to compare the peak floor acceleration(PFA)and FRS calculated from time history analyses(THA)with that generated using current seismic design codes and different methods in the literature.Major knowledge gaps are identified,including uncertainties associated with developing FRS,FRS generation methods for different types of buildings,the need for comprehensive studies on absolute acceleration,relative velocity,and relative displacement FRS,and the calibration of FRS by field observations during earthquakes.

Functional Analyses of Mammalian Reovirus Nonstructural Protein μNS

Genome replication of reovirus occurs in cytoplasmic inclusion bodies called viral factories or viroplasms. The viral nonstructural protein uNS, encoded by genome segment M3, is not a component of mature virions, but is expressed to high levels in infected cells and is concentrated in the infected cell factory matrix. Recent studies have demonstrated that uNS plays a central role in forming the matrix of these structures, as well as in recruiting other components to them for putative roles in genome replication and particle assembly.

Screening of Proteins Interacting with Nonstructural 1 Protein of H5N1 Avian Influenza Virus from T7-phage Display Library

Avian influenza virus（AIV） nonstructural 1（NS1） gene was amplified by real-time polymerse chain reac tion（RT-PCR） and inserted into pET28a, then transformed into E. coli BL21（DE3） competent cell. With the induction of isopropyl-β-D-thiogalactoside（IPTG） and the purification of Ni-NTA column, we finally obtained purified NS1 protein. T7-phage display system was used to screen the proteins that interacted with NS1 from lung cell cDNA li brary. The selected positive clones were identified by DNA sequencing and analyzed by BLAST program in Gene Bank. Two proteins were obtained as NS1 binding proteins, Homo sapiens nucleolar and coiled-body phosphoprotein 1（NOLC1） and Homo sapiens similar to colon cancer-associated antigen. By co-immunoprecipitation and other me thods, Homo sapiens NOLC1 was found to interact with the NS1 protein, the results would provide the basis for fur ther studying biological function of NS1 protein.

Identification of the strain-specifically truncated nonstructural protein 10 of porcine reproductive and respiratory syndrome virus in infected cells

The nonstructural protein 10（nsp10） of porcine reproductive and respiratory syndrome virus（PRRSV） encodes for helicase which plays a vital role in viral replication. In the present study, a truncated form of nsp10, termed nsp10 a, was found in PRRSV-infected cells and the production of nsp10 a was strain-specific. Mass spectrometric analysis and deletion mutagenesis indicated that nsp10 a may be short of about 70 amino acids in the N terminus of nsp10. Further studies by rescuing recombinant viruses showed that the Glu-69 in nsp10 was the key amino acid for nsp10 a production. Finally, we demonstrated that nsp10 a exerted little influence on the growth kinetics of PRRSV in vitro.

Effects of AG1 and AG2 QTLs on Nonstructural Carbohydrate and Seed Management Options for Rice Seedling Growth and Establishment under Flooding Stress

Rice(Oryza sativa)plants acquired excess photosynthates in the form of nonstructural carbohydrates(NSCs)in their stems and grain.Despite keen interest in rice NSC,the dynamics of NSC accumulation,translocation and re-accumulation have not yet been well investigated.AG1 and AG2 QTLs associated with flooding tolerance through catalyzing starch into soluble sugar in germinating seeds.Here we conducted three experiments,greenhouse and field to lay the groundwork for large-scale diversity studies on grain NSC and some agronomic traits under direct-seeded rice(DSR)system,using elite lines incorporating AG1,AG2 and AG1-AG2 QTLs into the popular varieties PSB Rc82 and Ciherang-Sub1 along with the donors Kho Hlan On(AG1)and Ma-Zhan Red(AG2).In germinating seedlings,soluble sugars increased,while starch concentration decreased gradually especially in the tolerant checks and AG1-AG2 introgression lines under flooded soil.Soluble sugar accumulation in stem started to increase from the vegetative stage and peaked at the panicle initiation stage then gradually decreased towards the maturity stage.But Sub1-AG lines had higher sugar and starch concentrations at different growth stages than other genotypes in wet season 2016 and dry season 2017.Plant survival rate was positively correlated with the stem NSC at the early vegetative stage(21 days after sowing),and stem NSC was positively associated with plant height at different growth stages.Among the tested seeding rate,the most suitable seeding rate,4 g/m2 with shallow burial depth(0.5 cm),resulted in better seedling establishment,relatively higher seedling vigor index and higher leaf area index under flooding in DSR system.Introgression of AG1-AG2 QTLs had no any negative impact on nonstructural carbohydrate,germination rate,and growth and biomass production.

Nuclear localization of Sindbis virus nonstructural protein nsP2

In early infection, approximately 10% of nonstruc-tural protein nsP2 of Sindbis virus was transported into the nuclei of virus-infected BHK-21 cells. Nuclear nsP2 was dominantly associated with nuclear matrix. During the course of infection, increasing amounts of nsP2 accumulated in the nuclear fraction. A prominent accumulation of nuclear nsP2 occurred early in infection, from 1 h to 3 h postinfection. Meanwhile, a weak NTPase activity was found to be associated with the immunocomplexed nsP2. Nuclear localization of nsP2 and its possible role were discussed in relation to the inhibition of host macro-molecular synthesis.

Hepatitis C virus nonstructural protein NS_3 and telomerase activity

OBJECTIVE: To study the effect of hepatitis C virus nonstructural protein NS(3) (HCV NS3) on telomerase activity and carcinogenesis. METHODS: Streptavidin-peroxidase (SP) conjugated method was used to detect the expression of HCV NS(3) protein in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' and pRcHCNS(3)-3'. Telomerase activity was detected by an in situ telomerase activity labeling method, telomeric repeat amplification protocol polymerase chain reaction (TRAP-PCR) and telomerase PCR enzyme linked immunosorbent assay (ELISA) technology in the transfected and non-transfected NIH3T3 cells. RESULTS: HCV NS(3) protein was expressed in the NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' expressing HCV NS(3) C-terminal deleted protein or with plasmid pRcHCNS(3)-3' expressing HCV NS(3) N-terminal deleted protein. The positive signal of HCV NS(3) protein was localized in the cytoplasm of NIH3T3 cells, and the signal intensity of the former was stronger. Telomerase activity in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' was stronger than that in NIH3T3 cells transfected with plasmid pRcHCNS(3)-3' (P