Overexpression of the Watermelon Ethylene Response Factor ClERF069 in Transgenic Tomato Resulted in Delayed Fruit Ripening

Watermelon fruit undergoes distinct development stages with dramatic changes during fruit ripening.To date,the molecular mechanics of watermelon ripening remain unclear.Genetic and transcriptome evidences suggested that the ethylene response factor(ERF)gene ClERF069 may be an important candidate factor affecting watermelon fruit ripening.To dissect the roles of ClERF069 in fruit ripening,structure and phylogenetic analysis were performed using the amplified full-length sequence.Normal-ripening watermelon 97103,non-ripening watermelon PI296341-FR and the RIL population were used to analyze ClERF069 expression dynamics and the correlation with fruit ripening indexs.The results indicated that ClERF069 belongs to ERF family group VI and show high homology(83%identity)to melon ERF069-like protein.ClERF069 expression in watermelon flesh was negatively correlated with fruit lycopene content and sugar content during fruit ripening progress.Further transgenic evidences indicated that overexpression of 35S:ClERF069 in tomato noticeably delayed the ripening process up to 5.2 days.Lycopene,β-carotenoid accumulation patterns were altered and ethylene production patterns in transgenic fruits was significantly delayed during fruit ripening.Taken together,watermelon ethylene response factor ClERF069 was concluded to be a negative regulator of fruit ripening.

The Openness Degree Study of the Jiaoshiba Shale Gas, Sichuan Basin, China–Potential Factor Responsible for Reversed Isotope Series

Objective Reversed alkane δ;C and δ;H values in many prolific shale plays all over the world have aroused much attention in the study of the formation mechanism of reversed isotope series in alkanes in the past few years（Zou Caineng et al., 2016）. Although many researchers have put forward different hypotheses, the mechanism has not been well understood yet. The openness degree of oil and gas system

Serum response factor:Look into the gut

Serum response factor(SRF) is a transcription factor that regulates many genes involved in cellular activities such as proliferation,migration,differentiation,angiogenesis,and apoptosis.Although it has only been known for about two decades,SRF has been studied extensively.To date,over a thousand SRF studies have been published,but it still remains a hot topic.Due to its critical role in mesoderm-derived tissues,most of the SRF studies focused on muscle structure/function,cardiovascular development/maintenance,and smooth muscle generation/repair.Recently,SRF has received more attention in the digestive field and several important discoveries have been made.This review will summarize what we have learned about SRF in the gastrointestinal tract and provide insights into possible future directions in this area.

Poly(ADP-ribosyl)ation of Apoptosis Antagonizing Transcription Factor Involved in Hydroquinone-Induced DNA Damage Response

The molecular mechanism of DNA damage induced by hydroquinone （HQ） remains unclear. Poly（ADP-ribose） polymerase-1 （PARP-1） usually works as a DNA damage sensor, and hence, it is possible that PARP-1 is involved in the DNA damage response induced by HQ. In TK6 cells treated with HQ, PARP activity as well as the expression of apoptosis antagonizing transcription factor （AATF）, PARP-1, and phosphorylated H2AX （v-H2AX） were maximum at 0.5 h, 6 h, 3 h, and 3 h, respectively. To explore the detailed mechanisms underlying the prompt DNA repair reaction, the above indicators were investigated in PARP-l-silenced cells. PARP activity and expression of AATF and PARP-1 decreased to 36%, 32%, and 33%, respectively, in the cells; however, y-H2AX expression increased to 265%. Co-immunoprecipitation （co-IP） assays were employed to determine whether PARP-1 and AATF formed protein complexes. The interaction between these proteins together with the results from IP assays and confocal microscopy indicated that poly（ADP-ribosyl）ation {PARylation） regulated AATF expression, in conclusion, PARP-1 was involved in the DNA damage repair induced by HQ via increasing the accumulation of AATF through PARylation.

Response reduction factor of irregular RC buildings in Kathmandu valley

Most current seismic design includes the nonlinear response of a structure through a response reduction factor （R）. This allows the designer to use a linear elastic force-based approach while accounting for nonlinear behavior and deformation limits. In fact, the response reduction factor is used in modem seismic codes to scale down the elastic response of a structure. This study focuses on estimating the actual ＇R＇ value for engineered design/construction of reinforced concrete （RC） buildings in Kathmandu valley. The ductility and overstrength of representative RC buildings in Kathmandu are investigated. Nonlinear pushover analysis was performed on structural models in order to evaluate the seismic performance of buildings. Twelve representative engineered irregular buildings with a variety of characteristics located in the Kathmandu valley were selected and studied. Furthermore, the effects of overstrength on the ductility factor, beam column capacity ratio on the building ductility, and load path on the response reduction factor, are examined. Finally, the results are further analyzed and compared with different structural parameters of the buildings.

Serum response factor promotes axon regeneration following spinal cord transection injury

Studies have snown that serum response factor is beneficaial for axonar regeneration of peripheral herves.However,Its role after central nervous system injury remains unclear. In this study,we established a rat model of T9-T10 spinal cord transection injury.We found that the expression of serum response factor in injured spinal cord gray matter neurons gradually increased with time,reached its peak on the 7^(th) day,and then gradually decreased.To investigate the role of serum response factor,we used lentivirus vecto rs to ove rexpress and silence serum response factor in spinal cord tissue.We found that overexpression of serum response factor promoted motor function recovery in rats with spinal cord injury.Qualitative observation of biotinylated dextran amine anterograde tra cing showed that ove rexpression of serum response factor increased nerve fibers in the injured spinal co rd.Additionally,transmission electron microscopy showed that axon and myelin sheath morphology was restored.Silencing serum response factor had the opposite effects of ove rexpression.These findings suggest that serum response factor plays a role in the recovery of motor function after spinal cord injury.The underlying mechanism may be related to the regulation of axonal regeneration.

Assessment of seismic design response factors of concrete wall buildings

To verify the seismic design response factors of high-rise buildings, five reference structures, varying in height from 20- to 60-stories, were selected and designed according to modern design codes to represent a wide range of concrete wall structures. Verified fiber-based analytical models for inelastic simulation were developed, considering the geometric nonlinearity and material inelasticity of the structural members. The ground motion uncertainty was accounted for by employing 20 earthquake records representing two seismic scenarios, consistent with the latest understanding of the tectonic setting and seismicity of the selected reference region （UAE）. A large number of Inelastic Pushover Analyses （IPAs） and Incremental Dynamic Collapse Analyses （IDCAs） were deployed for the reference structures to estimate the seismic design response factors. It is concluded that the factors adopted by the design code are adequately conservative. The results of this systematic assessment of seismic design response factors apply to a wide variety of contemporary concrete wall buildings with various characteristics.

Light Promotes Protein Stability of Auxin Response Factor 7#

Light is an environmental signaling,whereas Aux/IAA proteins and Auxin Response Factors(ARFs)are regulators of auxin signalling.Aux/IAA proteins are unstable,and their degradation dependents on 26S ubiquitin-proteasome and is promoted by Auxin.Auxin binds directly to a SCF-type ubiquitin-protein ligase,TIR1,facilitates the interaction between Aux/IAA proteins and TIR1,and then the degradation of Aux/IAA proteins.A few studies have reported that some ARFs are also unstable proteins,and their degradation is also mediated by 26S proteasome.In this study,by using of antibodies recognizing endogenous ARF7 proteins,we found that protein stability of ARF7 was affected by light.By expressing MYC tagged ARF activators in protoplasts,we found that degradation of ARF7 was inhibited by 26 proteasome inhibitors.In addition,at least ARF5 and ARF19 were also unstable proteins,and degradation of ARF5 via 26S proteasome was further confirmed by using stable transformed plants overexpressing ARF5 with a GUS tag.

Predictive active control of building structures using LQR and artificial intelligence

This study presents a neural network-based model for predicting linear quadratic regulator(LQR)weighting matrices for achieving a target response reduction.Based on the expected weighting matrices,the LQR algorithm is used to determine the various responses of the structure.The responses are determined by numerically analyzing the governing equation of motion using the state-space approach.For training a neural network,four input parameters are considered:the time history of the ground motion,the percentage reduction in lateral displacement,lateral velocity,and lateral acceleration,Output parameters are LQR weighting matrices.To study the effectiveness of an LQR-based neural network(LQRNN),the actual percentage reduction in the responses obtained from using LQRNN is compared with the target percentage reductions.Furthermore,to investigate the efficacy of an active control system using LQRNN,the controlled responses of a system are compared to the corresponding uncontrolled responses.The trained neural network effectively predicts weighting parameters that can provide a percentage reduction in displacement,velocity,and acceleration close to the target percentage reduction.Based on the simulation study,it can be concluded that significant response reductions are observed in the active-controlled system using LQRNN.Moreover,the LQRNN algorithm can replace conventional LQR control with the use of an active control system.

Improvement of Copper-inducible Gene Expression System for Plant

The copper-regulated gene expression system has been developed to control spacial and temporal expression of transgene in plant. It comprises two parts: (1) ace I gene encoding copper-responsive transcription factor under the control of a constitutive or organ-specific promoter, and (2) a gene of interest under the control of a chimeric promoter consisting of the CaMV 35S (-90 to +8) promoter linked to the metal responsive element (MRE) carrying activating copper-metallothionein expression (ACE1)-binding sites. Here, the effectiveness of two different ACE1-binding cis -elements which derive from 5'-regulatory region of yeast metallothionein gene was investigated in transgenic tobacco (Nicotiana tabacum L. cv. W38). The results revealed that the MRE (-210 to -126) could increase the system inducibility by 50% - 100% compared with the previously reported MRE (-148 to -105). It is potential to use the copper-inducible system to control valuable gene traits in plant biotechnology.

Plant fertility defects induced by the enhanced expression of microRNA167

The plant hormone auxin plays a critical role in regulating plant growth and development. Recent advances have been made in the understanding of auxin response pathways, primarily by the characterization of auxin response mutants in Arabidopsis. In addition, microRNAs （miRNAs） have been shown to be critical regulators of genes important for normal plant development and physiology. However, little is known about possible interactions between miRNAs and hormonal signaling during normal development. Here we show that an Arabidopsis microRNA, miR167, which has a complementary sequence to a portion of the A UXINRESPONSE FACTOR6 （ARF6） and ARF8 mRNAs, can cause transcript degradation for ARF8, but not for ARF6. We report phenotypic characterizations of 35S：：MIR167b transgenic lines, and show that severe 35S：：MIR167b transgenic lines had phenotypes similar to those of an arf6 arf8 double mutant. The transgenic phenotypes suggest that miR167 may repress ARF6 at the level of translation. We demonstrate that the transgenic plants are defective in all four whods of floral organs. In the transgenic flowers, filaments were abnormally short, anthers could not properly release pollen, and pollen grains did not germinate. Our results provide an important link between the miRNA-mediated regulatory pathway of gene expression and the auxin signaling network promoting plant reproductive development.

Identification and characterization of MeERF genes and their targets in pathogen response by cassava(Manihot esculenta)

Cassava,Manihot esculenta Crantz (Me),is a major dietary source of calories for over 700 million people in tropical regions.The production of cassava is constantly threatened by cassava bacterial blight (CBB),caused by Xanthomonas axonopodis pv.manihotis (Xam).The gene resources for CBB-resistant breeding of cassava are limited.In model plant species,ethylene response factors play important roles in response to pathogen infection.In this study,cassava ethylene response factors (MeERFs) were identified and characterized as the first step in studying their potential for CBB-resistant breeding of cassava.In the cassava genome 155 MeERFs were identified,of which 23 were induced by Xam infection.The promoter regions of204 genes harbored GCC-box that had the potential to interact with MeERFs.Using 37 transcriptomes derived from Xam infection treatment,four gene co-expression modules for the MeERFs and GCC-box containing genes were constructed.Six MeERFs were associated with two GCC-box containing genes:transcription initiation factor TFIIE subunit beta (MeTFIIE),and histone-lysine N-methyltransferase ASHR1 (MeASHR1).Dual-luciferase reporter assays showed that MeERF10 and MeERF58 positively regulated Me TFIIE;MeERF137 negatively regulated Me TFIIE;MeERF10 and MeERF137 positively regulated Me ASHR1;and MeERF35 negatively regulated Me ASHR1.The four MeERFs may mediate pathogen response by regulating the expression of the two GCC-box containing genes.

COMPOSITION DETERMINATION OF BINARY POLYMER MIXTURES BY SIZE EXCLUSION CHROMATOGRAPHY WITH LIGHT SCATTERING DETECTION

Base on the principle of absolute quantification of size exclusion chromatography （SEC）, a light scattering （LS） detector coupled with a concentration detector （refractive index detector） is utilized to determine the compositions of complicated binary mixtures. A theoretical analysis predicts that the response factors for both LS and RI detectors are linear functions with the composition of any specified polymer mixtures in the binary polymer mixtures. Two pairs of complicated binary mixtures were used to test the theory mentioned in the present paper, and the experimental results show an excellent accordance with the theory.

Anti-inflammatory properties of lipoxin A4 protect against diabetes mellitus complicated by focal cerebral ischemia/reperfusion injury

Lipoxin A4 can alleviate cerebral ischemia/reperfusion injury by reducing the inflammatory reaction,but it is currently unclear whether it has a protective effect on diabetes mellitus complicated by focal cerebral ischemia/reperfusion injury.In this study,we established rat models of diabetes mellitus using an intraperitoneal injection of streptozotocin.We then induced focal cerebral ischemia/reperfusion injury by occlusion of the middle cerebral artery for 2 hours and reperfusion for 24 hours.After administration of lipoxin A4 via the lateral ventricle,infarction volume was reduced,the expression levels of pro-inflammatory factors tumor necrosis factor alpha and nuclear factor-kappa B in the cerebral cortex were decreased,and neurological functioning was improved.These findings suggest that lipoxin A4 has strong neuroprotective effects in diabetes mellitus complicated by focal cerebral ischemia/reperfusion injury and that the underlying mechanism is related to the anti-inflammatory action of lipoxin A4.

Quantitative Analysis Using Fourier Transform Ion Cyclotron Resonance Mass Spectrometry and Correlation between Mass Spectrometry Data and Sulfur Content of Crude Oils

Fourier transform ion cyclotron resonance mass spectrometry(FT-ICR MS) has become a powerful tool for analyzing the detailed composition of petroleum samples. However, the correlation between the numerous peaks obtained by FT-ICR MS and bulk properties of petroleum samples is still a challenge. In this study, the internal standard method was applied for the quantitative analysis of four straight-run vacuum gas oils(VGO) by atmospheric pressure photoionization(APPI) FT-ICR MS. The heteroatom class distribution of these VGO samples turned to be different when the concentration changed. Linear relationship between the normalized abundance and the concentration of VGO samples was identified for the total aromatic compounds, aromatic hydrocarbons, S1 and N1 species. The differences of the response factors were also discussed. The sulfur contents of a series of crude oils were proved to be linear with the FT-ICR MS data calibrated by the response factor of S1 species. This study demonstrated the feasibility of the internal standard method in quantitative analysis with APPI FT-ICR MS, and the bulk properties of petroleum samples could be correlated directly with the FT-ICR MS data.

Efficient Anomalous Forcings for Linear Problems

For linear forcing problems, a method is developed to provide a set of forcing modes which form a complete orthonormal basis for the finite-time response to steady forcing in the energy inner product space. The forcing modes are found by calculating eigenvectors of a positive definite and symmetric matrix determined from given equations of motion. The amplitude of responses to forcing modes is given in terms of the associated eigenvalues. This method is used in a nondivergent barotropic model linearized about the 300 hPa zonally-varying climatological flow both for northern summertime and wintertime. The results show that the amplitude of response varies considerably with different forcing modes. Only a few of forcing modes associated with the leading eigenvalues, called efficient forcing mode, can excite significant response. The efficient forcing modes possess highly localized spatial structure with wavetrain appearance. Most of the efficient forcings are located to the south of regions of the jet cores. The forcings located over polar regions are also efficient. In addition, the response is larger in wintertime than in summertime for a given forcing.

Extreme value of wind-excited response considering influence of bandwidth

This paper addresses the peak factors of wind- excited responses including alongwind, acrosswind tall building responses and vortex-induced vibration considering the bandwidth parameter. The influence of bandwidth parameter on the peak factor is investigated using advanced upcrossing theory taking the bandwidth influence into account. Results show that Davenport＇s formula without consideration of bandwidth parameter servers well in general. However, the advanced upcrossing theory leads to a better prediction of the peak factor of wind-induced response of very lightly damped buildings.

AtDREB2A-CA Influences Root Architecture and Increases Drought Tolerance in Transgenic Cotton

Drought is a major environmental factor limiting cotton (Gossypium hirsutum L.) productivity worldwide and projected climate changes could increase their negative effects in the future. Thus, targeting the molecular mechanisms correlated with drought tolerance without reducing productivity is a challenge for plant breeding. In this way, we evaluated the effects of water deficit progress on AtDREB2A-CA transgenic cotton plant responses, driven by the stress-inducible rd29 promoter. Besides shoot and root morphometric traits, gas exchange and osmotic adjustment analyses were also included. Here, we present how altered root traits shown by transgenic plants impacted on physiological acclimation responses when submitted to severe water stress. The integration of AtDREB2A-CA into the cotton genome increased total root volume, surface area and total root length, without negatively affecting shoot morphometric growth parameters and nor phenotypic evaluated traits. Additionally, when compared to wild-type plants, transgenic plants (17-T0 plants and its progeny) highlighted a gradual pattern of phenotypic plasticity tosome photosynthetic parameters such as photosynthetic rate and stomatal conductance with water deficit progress. Transgene also promoted greater shoot development and root robustness (greater and deeper root mass) allowing roots to grow into deeper soil layers. The same morpho-physiological trend was observed in the subsequent generation (17.6-T2). Our results suggest that the altered root traits shown by transgenic plants are the major contributors to higher tolerance response, allowing the AtDRE2A-CA-cotton plants to maintain elevated stomatal conductance and assimilate rates and, consequently, reducing their metabolic costs involved in the antioxidant responses activation. These results also suggest that these morpho-physiological changes increased the number of reproductive structures retained per plant (26% higher) when compared with its non-transgenic counterpart. This is the first report of cotton plants overexpressing the AtDRE2A-CA transcription factor, demonstrating a morpho-physiological and yield advantages under drought stress, without displaying any yield penalty under irrigated conditions. The mechanisms by which the root traits influenced the acclimation of the transgenic plants to severe water deficit conditions are also discussed. These data present an opportunity to use this strategy in cotton breeding programs in order to improve drought adaptation toward better rooting features.

Cotton Major Latex Protein 28 Functions as a Positive Regulator of the Ethylene Responsive Factor 6 in Defense against Verticillium dahliae

In this study, we identified a defense-related major latex protein （MLP） from upland cotton （designated GhMLP28） and investigated its functional mechanism. GhMLP28 transcripts were ubiquitously present in cotton plants, with higher accumulation in the root. Expression of the GhMLP28 gene was induced by Verticillium dahliae inoculation and was responsive to defense signaling molecules, including ethylene, jas- monic acid, and salicylic acid. Knockdown of GhMLP28 expression by virus-induced gene silencing re- sulted in increased susceptibility of cotton plants to V. dahliae infection, while ectopic overexpression of GhMLP28 in tobacco improved the disease tolerance of the transgenic plants. Further analysis revealed that GhMLP28 interacted with cotton ethylene response factor 6 （GhERF6） and facilitated the binding of GhERF6 to GCC-box element. Transient expression assay demonstrated that GhMLP28 enhanced the tran- scription factor activity of GhERF6, which led to the augmented expression of some GCC-box genes. GhMLP28 proteins were located in both the nucleus and cytoplasm and their nuclear distribution was dependent on the presence of GhERF6. Collectively, these results demonstrate that GhMLP28 acts as a positive regulator of GhERF6, and synergetic actions of the two proteins may contribute substantially to protection against V. dahliae infection in cotton plants.

WRKY transcription factors in plant responses to stresses

The WRKY gene family is among the largest families of transcription factors （TFs） in higher plants. By regulating the plant hormone signal transduction pathway, these TFs play critical roles in some plant processes in response to biotic and abiotic stress, Various bodies of research have demonstrated the important biological functions of WRKY TFs in plant response to different kinds of biotic and abiotic stresses and working mecha- nisms. However, very li2ttle summarization has been done to review their research progress. Not iust important TFs function in plant response to biotic and abiotic stresses, WRKY also participates in carbohydrate synthesis, senes- cence, development, and secondary metabolites synthesis. WRKY proteins can bind to W-box （TGACC （A/T）） in the promoter of its target genes and activate or repress the expression of downstream genes to regulate their stress response. Moreover, WRKY proteins can interact with other TFs to regulate plant defensive responses. In the present review, we focus on the structural characteristics of WRKY TFs and the research progress on their functions in plant responses to a variety of stresses.