An evolutionary perspective on differential regulation of zinc and cadmium homeostatis genes in Arabidopsis thaliana and Noccaea caerulescens

Some plants can tolerate and accumulate unusually high levels of toxic metals, and the analysis of such plants can provide insights into the ecology of environments that are polluted with heavy metals due to human industrial activities. The study of heavy metal hyperaccumulators such as Noccaea caerulescens can show how plants cope with excess metals and increase their fitness when growing in metalliferous environments. In this thesis, I compared the molecular mechanisms of zinc (Zn) homeostasis and cadmium (Cd) response in the hyperaccumulator species N. caerulescens and its non-accumulator relative Arabidopsis thalianaby investigating the regulation of the ZNT1/ZIP4gene that promotes Zn uptake and Zn/Cd tolerance.I also studied the ecological advantages of metal hyperaccumulators in nature and determined the DNA sequences of the N. caerulescens transcriptometo find candidate genes that control metal hyperaccumulation and provide an evolutionary perspective on the emergence of this trait. The functional characterization of the N. caerulescens and A. thaliana Zn-transporter genes NcZNT1 and AtZIP4 (and their promoters) showed how their differential expression pattern contributed to their role in metal tolerance and accumulation. The NcZNT1 gene is induced by Zn deficiency and the NcZNT1 protein is localized in the plasma membrane. Transgenic N. caerulescens roots containing a transgene for the green fluorescent protein (GFP) driven by the NcZNT1promoter revealed GFP fluorescence localized to pericycle and vascular tissues. This suggests that NcZNT1 contributes to metal loading into the xylem and long-distance metal transport. The overexpression of NcZNT1 in A. thaliana increased Zn and Cd tolerance and the capacity to accumulate these metals compared to wild-type plants. These results suggest that NcZNT1 plays an important role in Zn and Cd hypertolerance and hyperaccumulation in N. caerulescens, where it is expressed in both Zn-sufficient and Zn-excess conditions. The differential activity of the NcZNT1 and AtZIP4 promoters in N. caerulescens and A. thaliana implies that different cis-regulatory elements and trans-regulatory factors are present in both species. The ecological advantage of hyperaccumulators in metal-contaminated soils was investigated by studying a natural nas1mutant, in which the Nicotianamine Synthase1 gene is disrupted by a transposon insertion. This mutant allele was found in three natural N. caerulescens populations, which were compared to wild-type plants in terms of phenotype and adaptive advantage. Although the transposon disrupted the gene, the loss of NAS1 gene activity was compensated by enhanced expression of NAS3 and NAS4, resulting in increased nicotianamine (NA) production, which enhanced the Zn and Cd accumulation in the nas1 mutants. This increased their metal sensitivity compared to wild-type plants, but also made them more toxic towards Pieris rapae caterpillars, which developed more slowly and gained less weight when fed on mutant plants exposed to excess Zn or Cd. Therefore, the possible selective advantage of the nas1 mutant in nature is high metal accumulation and the protection of plants from herbivores. Differences in nas1 allele frequency among the three natural populations suggests that nas1 alleles experience different degrees of natural selection or may be at different stages on the route to fixation. Molecular evolutionary studies involve the identification of candidate genes that play a role in adaptation. Therefore, a comprehensive set of transcript sequences was obtained from N. caerulescens accession Ganges (GA) by 454 pyrosequencing. In total, the collected 23,836 isotigs (putative transcripts) were grouped into 20,378 isogroups and 93.2% of them could be matched to Brassicaceae protein sequences, which allowedtheir functional annotation. A total of 87 isogroups was annotated as metal homeostasis related genes, including metal transporter families, metal chelator biosynthesis families, and metal tolerance gene families, which are candidate genes for the molecular analysis of heavy metal homeostasis mechanisms. A group of genes required for the synthesis of glucosinolates, which are important secondary metabolites that protect plants against herbivores, were also shown to be expressed in N. caerulescens. The METALLOTHIONEIN3 (MT3) gene was found to have been duplicated in the N. caerulescens genome, when compared to related Brassicaceae. These transcript sequences will provide an important tool to annotate the genome sequence of the N. caerulescensGanges accession, which is in progress. This genome sequence will also be the basis of genome comparisons between the different N. caerulescens accessions that have different levels of metal accumulation and metal tolerance and which may therefore differ in terms of gene expression levels or modes of actions that affect metal homeostasis.</p

Modeling asphalt pavement overlay transverse cracks using the genetic operation tree and Levenberg-Marquardt Method

[[abstract]]The Artificial Neural Network (ANN) and the nonlinear regression method are commonly used to build models from experimental data. However, the ANN has been criticized for incapable of providing clear relationships and physical meanings, and is usually regarded as a black box. The nonlinear regression method needs predefined and correct formula structures to process parameter search in terms of the minimal sum of square errors. Unfortunately, the formula structures of these models are often unclear and cannot be defined in advance. To overcome these challenges, this study proposes a novel approach, called ââLMGOT,ââ that integrates two optimization techniques: the LevenbergâMarquardt (LM) Method and the genetic operation tree (GOT). The GOT borrows the concept from the genetic algorithm, a famous algorithm for solving discrete optimization problems, to generate operation trees (OTs), which represent the structures of the formulas. Meanwhile, the LM takes advantage of its merit for solving nonlinear continuous optimization problems, and determines the coefficients in the GOTs that best fit the experimental data. This paper uses the LMGOT to investigate the data sets of pavement cracks from a 15-year experiment conducted by the Texas Departments of Transportation. Results show a concise formula for predicting the length of pavement transverse cracking, and indicate that the LMGOT is an efficient approach to building an accurate crack model.[[incitationindex]]SCI[[booktype]]紙

ARCHERY TRAINING IMPROVE POSTURAL CONTROL IN YOUNG CHILDREN

Exercise training might be beneficial for postural control (PC) during exercise and daily activities. The purpose of the study was to examine the effects of an eight-week archery training program on PC in young children during the standard, aiming, and archery shooting standing posture

Kinetic Parameters Evaluation for Isoprene Mixed with Aluminum Oxide

AbstractWe analyzed the basic hazard characteristics for isoprene in the manufacturing process by evaluating the runaway reaction, kinetic parameters and safety parameters in this study. Experiments were carried out by calculating the thermal decomposition reactions for isoprene and mixed with aluminum oxide in non-isothermal conditions by differential scanning calorimetry (DSC). Afterwards results indicated that isoprene had a conjugated double bond of unstable structure, and therefore it was prone to produce exothermic reaction during the process of polymerization. Aluminum oxide was applied to mix with isoprene, which could increase the heat release rate. Based on the results, safety information should be provided to government and relevant industries for prevention the accident occur in relevant plants

Hydrophobic Modification of Layered Clays and Compatibility for Epoxy Nanocomposites

Recent studies on the intercalation and exfoliation of layered clays with polymeric intercalating agents involving poly(oxypropylene)-amines and the particular uses for epoxy nanocomposites are reviewed. For intercalation, counter-ionic exchange reactions of clays including cationic layered silicates and anionic Al-Mg layered double hydroxide (LDH) with polymeric organic ions afforded organoclays led to spatial interlayer expansion from 12 to 92 angstrom (X-ray diffraction) as well as hydrophobic property. The inorganic clays of layered structure could be modified by the poly(oxypropylene) amine-salts as the intercalating agents with molecular weights ranging from 230 to 5,000 g/mol. Furthermore, natural montmorillonite (MMT) clay could be exfoliated into thin layer silicate platelets (ca. 1 nm thickness) in one step by using polymeric types of exfoliating agents. Different lateral dimensions of MMT, synthetic fluorinated Mica and LDH clays had been cured into epoxy nanocomposites. The hydrophobic amine-salt modification resulting in high spacing of layered or exfoliation of individual clay platelets is the most important factor for gaining significant improvements of properties. In particular, these modified clays were reported to gain significant improvements such as reduced coefficient of thermal expansion (CTE), enhanced thermal stability, and hardness. The utilization of these layered clays for initiating the epoxy self-polymerization was also reported to have a unique compatibility between clay and organic resin matrix. However, the matrix domain lacks of covalently bonded crosslink and leads to the isolation of powder material. It is generally concluded that the hydrophobic expansion of the clay inter-gallery spacing is the crucial step for enhancing the compatibility and the ultimate preparation of the advanced epoxy materials

Development of a multiplex event-specific PCR assay for detection of genetically modified rice

Global rice supplies have been found contaminated with unapproved varieties of genetically modified (GM) rice in recent years, which has led to product recalls in several of countries. Faster and more effective detection of GM contamination can prevent adulterated food, feed and seed from being consumed and grown, minimize the potential environmental, health or economic damage. In this study, a simple, reliable and cost-effective multiplex polymerase chain reaction (PCR) assay for identifying genetic modifications of TT51-1, Kemingdao1 (KMD1) and Kefeng6 (KF6) rice was developed by using the event-specific fragment. The limit of detection (LOD) for each event in the multiplex PCR is approximately 0.1%. Developed multiplex PCR assays can provide a rapid and simultaneous detection of GM rice

Submergence of the Sidebands in the Photon-assisted Tunneling through a Quantum Dot Weakly Coupled to Luttinger Liquid Leads

We study theoretically the photon-assisted tunneling through a quantum dot weakly coupled to Luttinger liquids (LL) leads, and find that the zero bias dc conductance is strongly affected by the interactions in the LL leads. In comparison with the system with Fermi liquid (FL) leads, the sideband peaks of the dc conductance become blurring for 1/2<g<1, and finally merge into the central peak for g<1/2, (g is the interaction parameter in the LL leads). The sidebands are suppressed for LL leads with Coulomb interactions strong enough, and the conductance always appears as a single peak for any strength and frequency of the external time-dependent field. Furthermore, the quenching effect of the central peak for the FL case does not exist for g<1/2.Comment: 9 pages, 4 figure

NTMG (N-terminal Truncated Mutants Generator for cDNA): an automatic multiplex PCR assays design for generating various N-terminal truncated cDNA mutants

The sequential deletion method is generally used to locate the functional domain of a protein. With this method, in order to find the various N-terminal truncated mutants, researchers have to investigate the ATG-like codons, to design various multiplex polymerase chain reaction (PCR) forward primers and to do several PCR experiments. This web server (N-terminal Truncated Mutants Generator for cDNA) will automatically generate groups of forward PCR primers and the corresponding reverse PCR primers that can be used in a single batch of a multiplex PCR experiment to extract the various N-terminal truncated mutants. This saves much time and money for those who use the sequential deletion method in their research. This server is available at http://oblab.cs.nchu.edu.tw:8080/WebSDL/

Indenopyridine hydrochloride induced testicular spermatogenesis failure with high seminal alkaline phosphatase levels in male dog

Indenopyridine hydrochloride (IH), an antispermatogenic agent, was tested to determine the testicular pathological changes, seminal spermatozoa concentrations and seminal plasma alkaline phosphatase levels in male dogs. A single oral dosage of 30 mg IH/kg BW induced the dissociation and premature release of germ cells into the lumens of seminiferous tubules. Ring-shaped spermatid nuclei, nuclear pykonosis of spermatocytes and multinucleated cell associations were also observed. Thereafter, the spermatogenic index (SI) significantly decreased one day after IH administration. Moreover, seminal spermatozoa concentrations decreased two weeks after drug treatment; and there was a statistically significant difference in spermatozoa production inhibited by IH compared to the control. Reversible spermatogenesis was noted 7 weeks after IH treatment in male dogs. Meanwhile, seminal plasma alkaline phosphatase levels also significantly increased two weeks after IH treatment. These data confirm that IH might induce a two-month inhibition of spermatogenesis in male dogs

Measuring CP violation and mass ordering in joint long baseline experiments with superbeams

We propose to measure the CP phase $\delta_{\rm CP}$, the magnitude of the neutrino mixing matrix element $|U_{e3}|$ and the sign of the atmopheric scale mass--squared difference $\Delta{\rm m}^2_{31}$ with a superbeam by the joint analysis of two different long baseline neutrino oscillation experiments. One is a long baseline experiment (LBL) at 300 km and the other is a very long baseline (VLBL) experiment at 2100 km. We take the neutrino source to be the approved high intensity proton synchrotron, HIPA. The neutrino beam for the LBL is the 2-degree off-axis superbeam and for the VLBL, a narrow band superbeam. Taking into account all possible errors, we evaluate the event rates required and the sensitivities that can be attained for the determination of $\delta_{\rm CP}$ and the sign of $\Delta m^2_{31}$. We arrive at a representative scenario for a reasonably precise probe of this part of the neutrino physics.Comment: 25 RevTEX pages, 16 PS figures, revised figure captions and references adde