Membrane Proteomics to Understand Enhancement Effects of Millimeter-Wave Irradiation on Wheat Root under Flooding Stress.

Millimeter-wave irradiation of wheat seeds enhances the growth of roots under flooding stress, but its mechanism is not clearly understood. To understand the role of millimeter-wave irradiation on root-growth enhancement, membrane proteomics was performed. Membrane fractions purified from wheat roots were evaluated for purity. H+-ATPase and calnexin, which are protein markers for membrane-purification efficiency, were enriched in a membrane fraction. A principal-component analysis of the proteomic results indicated that the millimeter-wave irradiation of seeds affects membrane proteins in grown roots. Proteins identified using proteomic analysis were confirmed using immunoblot or polymerase chain reaction analyses. The abundance of cellulose synthetase, which is a plasma-membrane protein, decreased under flooding stress; however, it increased with millimeter-wave irradiation. On the other hand, the abundance of calnexin and V-ATPase, which are proteins in the endoplasmic reticulum and vacuolar, increased under flooding stress; however, it decreased with millimeter-wave irradiation. Furthermore, NADH dehydrogenase, which is found in mitochondria membranes, was upregulated due to flooding stress but downregulated following millimeter-wave irradiation even under flooding stress. The ATP content showed a similar trend toward change in NADH dehydrogenase expression. These results suggest that millimeter-wave irradiation improves the root growth of wheat via the transitions of proteins in the plasma membrane, endoplasmic reticulum, vacuolar, and mitochondria.

Proteomic and Biochemical Approaches Elucidate the Role of Millimeter-Wave Irradiation in Wheat Growth under Flooding Stress.

Flooding impairs wheat growth and considerably affects yield productivity worldwide. On the other hand, irradiation with millimeter waves enhanced the growth of chickpea and soybean under flooding stress. In the current work, millimeter-wave irradiation notably enhanced wheat growth, even under flooding stress. To explore the protective mechanisms of millimeter-wave irradiation on wheat under flooding, quantitative proteomics was performed. According to functional categorization, proteins whose abundances were changed significantly with and without irradiation under flooding stress were correlated to glycolysis, reactive-oxygen species scavenging, cell organization, and hormonal metabolism. Immunoblot analysis confirmed that fructose-bisphosphate aldolase and ß tubulin accumulated in root and leaf under flooding; however, even in such condition, their accumulations were recovered to the control level in irradiated wheat. The abundance of ascorbate peroxidase increased in leaf under flooding and recovered to the control level in irradiated wheat. Because the abundance of auxin-related proteins changed with millimeter-wave irradiation, auxin was applied to wheat under flooding, resulting in the application of auxin improving its growth, even in such condition. These results suggest that millimeter-wave irradiation on wheat seeds improves the recovery of plant growth from flooding via the regulation of glycolysis, reactive-oxygen species scavenging, and cell organization. Additionally, millimeter-wave irradiation could promote tolerance against flooding through the regulation of auxin contents in wheat.

Proteomic and Biological Analyses Reveal the Effect on Growth under Flooding Stress of Chickpea Irradiated with Millimeter Waves.

Chickpea cultivated on marginal lands in arid and semiarid tropics is one of the food legumes, and its growth is reduced by flooding stress. Millimeter-wave irradiation has influences on organisms, and it improves the growth of plants such as soybean. To reveal the dynamic effects of millimeter-wave irradiation on chickpea under flooding, gel- and label-free proteomic analysis was conducted. Millimeter-wave irradiation improved chickpea growth and its tolerance to flooding stress. According to functional categorization, oppositely changed proteins were correlated with photosynthesis, fermentation, and protein degradation. Immunoblot analysis confirmed that RuBisCO activase and large subunits decreased in leaves under flooding; however, they are recovered in irradiated chickpea even if it was in this condition. The activity and accumulation of alcohol dehydrogenase increased in roots under flooding; however, this followed the same pattern. Cell death was significantly increased and decreased by flooding on unirradiated and irradiated chickpeas, respectively. These findings suggest that irradiation with millimeter waves on chickpea seeds improves the recovery of plant growth through regulation of photosynthesis in leaves and fermentation in roots. Furthermore, millimeter-wave irradiation might promote chickpea tolerance under flooding via the regulation of cell death.

Evaluation of high-stability optical beats in laser chaos by plasmonic photomixing.

The stability of optical beats in a chaotically oscillating laser is compared to that of a free-running continuous-wave laser using a highly efficient plasmonic photomixer. Using a chaotically oscillating laser diode, stable optical beats are observed over an operation current range of 60-90 mA. The optical spectra are stable even with frequent mode hopping. In contrast, optical beats in a free-running continuous-wave laser are not stable compared to those of a chaotically oscillating laser, because of intermittent hopping of the laser modes. The high stability of chaotically oscillating lasers makes these lasers promising candidates for optical pump sources in terahertz time-domain spectroscopy systems.

Proteomic Analysis of Irradiation with Millimeter Waves on Soybean Growth under Flooding Conditions.

Improving soybean growth and tolerance under environmental stress is crucial for sustainable development. Millimeter waves are a radio-frequency band with a wavelength range of 1-10 mm that has dynamic effects on organisms. To investigate the potential effects of millimeter-waves irradiation on soybean seedlings, morphological and proteomic analyses were performed. Millimeter-waves irradiation improved the growth of roots/hypocotyl and the tolerance of soybean to flooding stress. Proteomic analysis indicated that the irradiated soybean seedlings recovered under oxidative stress during growth, whereas proteins related to glycolysis and ascorbate/glutathione metabolism were not affected. Immunoblot analysis confirmed the promotive effect of millimeter waves to glycolysis- and redox-related pathways under flooding conditions. Sugar metabolism was suppressed under flooding in unirradiated soybean seedlings, whereas it was activated in the irradiated ones, especially trehalose synthesis. These results suggest that millimeter-waves irradiation on soybean seeds promotes the recovery of soybean seedlings under oxidative stress, which positively regulates soybean growth through the regulation of glycolysis and redox related pathways.

Enhancing terahertz magnetic near field induced by a micro-split-ring resonator with a tapered waveguide.

Substantial enhancement of terahertz magnetic near field achieved by the combination of a tapered metallic waveguide and a micro-split-ring resonator is demonstrated. The magnetic near field is probed directly via the magneto-optic sampling with a Tb3Ga5O12 crystal. The incident terahertz wave with a half-cycle waveform is generated by using the pulse-front tilting method. The magnetic near field at the resonant frequency is enhanced by more than 30 times through the combination of the waveguide and the resonator. The peak amplitude of the magnetic field with a damped oscillation waveform in the time domain is up to 0.4 T. The resonant frequency can be tuned by adopting different resonator designs. The mechanism of the enhancement is analyzed by performing calculations based on the finite element method. The strong terahertz magnetic near field enables the excitation of large-amplitude spin dynamics and can be utilized for an ultrafast spin control.

Polarization-variable emitter for terahertz time-domain spectroscopy.

We report on the progress in the development of linear polarization-variable multi-electrode emitters for terahertz time-domain spectroscopy. The results on its microfabrication, the finite element method modeling of appropriate bias distribution between electrodes, the finite-difference time-domain simulated spectral output, and actual experimental testing are presented. The rotation of the emitted terahertz field with linear polarization on an angle multiple of 45° can be achieved by synchronized bias and single polarizer rotations.

Low temperature-grown GaAs carrier lifetime evaluation by double optical pump terahertz time-domain emission spectroscopy.

We present the use of a "double optical pump" technique in terahertz time-domain emission spectroscopy as an alternative method to investigate the lifetime of photo-excited carriers in semiconductors. Compared to the commonly employed optical pump-probe transient photo-reflectance, this non-contact and room temperature characterization technique allows relative ease in achieving optical alignment. The technique was implemented to evaluate the carrier lifetime in low temperature-grown gallium arsenide (LT-GaAs). The carrier lifetime values deduced from "double optical pump" THz emission decay curves show good agreement with data obtained from standard transient photo-reflectance measurements on the same LT-GaAs samples grown at 250 °C and 310 °C.

Cherenkov-phase-matched nonlinear optical detection and generation of terahertz radiation via GaAs with metal-coating.

Terahertz (THz) wave detection and emission via Cherenkov-phase-matched nonlinear optical effects at 1.55-µm optical wavelength were demonstrated using a GaAs with metal-coating (M-G-M) and bare GaAs as a reference sample in conjunction with a metallic tapered parallel-plate waveguide (TPPWG). The metal-coated GaAs is superior to the bare wafer both as a THz electro-optic detector and as an emitter. Significant improvements in the detection and emission efficiency were obtained by utilizing a metal-coating due to better confinement and lower loss of the THz waves propagating in the M-G-M compared with bare GaAs.

A matter of symmetry: terahertz polarization detection properties of a multi-contact photoconductive antenna evaluated by a response matrix analysis.

While terahertz time domain spectroscopy (THz-TDS) is a well-established technique, polarization sensitive measurements are challenging due to the need of broadband polarization devices. Here, we characterize our recently introduced multi-contact photoconductive detector antenna with a response matrix analysis. We show that the lead lines attached to electrodes reduce the antenna symmetry and thereby influence the properties of the response matrices. With a wire grid polarizer, we simulate a sample influencing the polarization angle and the intensity of the incident THz pulse. Evaluating the measurements with the response matrix analysis, our results show a well agreement of the adjusted and measured polarization angles and intensities over a frequency range from 0.25 to 0.8 THz.

Confined photocarrier transport in InAs pyramidal quantum dots via terahertz time-domain spectroscopy.

We present experimental demonstration of photocarrier dynamics in InAs quantum dots (QDs) via terahertz (THz) time-domain spectroscopy (TDS) using two excitation wavelengths and observing the magnetic field polarity characteristics of the THz signal. The InAs QDs was grown using standard Stranski-Krastanow technique on semi-insulating GaAs substrate. Excitation pump at 800 nm- and 910 nm-wavelength were used to distinguish THz emission from the InAs/GaAs matrix and InAs respectively. THz-TDS at 800 nm pump revealed intense THz emission comparable to a bulk p-InAs. For 910 nm pump, the THz emission generally weakened and upon applying external magnetic field of opposite polarities, the THz time-domain plot exhibited anomalous phase-shifting. This was attributed to the possible current-surge associated with the permanent dipole in the QD.

A polarization-sensitive 4-contact detector for terahertz time-domain spectroscopy.

A light polarization angle-sensitive photoconductive detector for terahertz time-domain spectroscopy is computer-modeled, microfabricated, and tested. The experimental results show good agreement with the linear angular response for an ideal detector. The detector's frequency, angular, and crosstalk responses are discussed in the context of theoretical and experimental considerations.

Millmeter-wave irradiation regulates mRNA-expression and the ubiquitin-proteasome system in wheat exposed to flooding stress.

The irradiation with millimeter-wave (MMW) of wheat seeds promotes root growth under flooding stress; however, its role is not completely clarified. Nuclear proteomics was performed, to reveal the role of MMW irradiation in enhancing flooding tolerance. The purity of nuclear fractions purified from roots was verified. Histone, which is a protein marker for nuclear-purification efficiency, was enriched; and cytosolic ascorbate peroxidase was reduced in the nuclear fraction. The principal-component analysis of proteome displayed that the irradiation of seeds affected nuclear proteins in roots grown under flooding stress. Proteins detected using proteomic analysis were verified using immunoblot analysis. Histone H3 accumulated under flooding stress; however, it decreased to the control level by irradiation. Whereas the ubiquitin accumulated in roots grown under stress when seeds were irradiated. These results suggest that MMW irradiation improves wheat-root growth under flooding stress through the regulation of mRNA-expression level and the ubiquitin-proteasome system. SIGNIFICANCE: To reveal the role of millimeter-wave irradiation in enhancing flooding tolerance in wheat, nuclear proteomics was performed. The principal-component analysis of proteome displayed that irradiation of seeds affected nuclear proteins in roots grown under flooding stress. Proteins detected using proteomic analysis were verified using immunoblot analysis. Histone H3 accumulated under flooding stress; however, it decreased to the control level with irradiation. Whereas the ubiquitin accumulated in roots grown under stress when seeds were irradiated. These results suggest that millimeter-wave irradiation improves wheat-root growth under flooding stress through the regulation of mRNA-expression level and the ubiquitin-proteasome system.

Non-ellipsometric detection of terahertz radiation using heterodyne EO sampling in the Cherenkov velocity matching scheme.

A new electro-optic (EO) sampling scheme, which we refer to as "heterodyne EO sampling", for detection of pulsed terahertz (THz) waves is proposed and experimentally demonstrated. In this heterodyne EO sampling scheme, the intensity change in the optical probe pulse induced by a THz field in a nonlinear crystal is measured without any polarization optics. Applied in combination with the non-collinear Cherenkov velocity matching technique, this method allows one to detect pulsed THz waves efficiently and easily using a simpler optical setup as compared to the conventional ellipsometric EO sampling method.

Terahertz emission from Indium Oxide films grown on MgO substrates using sub-bandgap photon energy excitation.

Indium oxide (In2O3) films grown by thermal oxidation on MgO substrates were optically excited by femtosecond laser pulses having photon energy lower than the In2O3 bandgap. Terahertz (THz) pulse emission was observed using time domain spectroscopy. Results show that THz emission saturates at an excitation fluence of ~400 nJ/cm2. Even as two-photon absorption has been excluded, the actual emission mechanism has yet to be confirmed but is currently attributed to carriers due to weak absorption from defect levels that are driven by a strain field at the interface of the substrate and the grown film.

Exploring Biomolecular Self-Assembly with Far-Infrared Radiation.

Physical engineering technology using far-infrared radiation has been gathering attention in chemical, biological, and material research fields. In particular, the high-power radiation at the terahertz region can give remarkable effects on biological materials distinct from a simple thermal treatment. Self-assembly of biological molecules such as amyloid proteins and cellulose fiber plays various roles in medical and biomaterials fields. A common characteristic of those biomolecular aggregates is a sheet-like fibrous structure that is rigid and insoluble in water, and it is often hard to manipulate the stacking conformation without heating, organic solvents, or chemical reagents. We discovered that those fibrous formats can be conformationally regulated by means of intense far-infrared radiations from a free-electron laser and gyrotron. In this review, we would like to show the latest and the past studies on the effects of far-infrared radiation on the fibrous biomaterials and to suggest the potential use of the far-infrared radiation for regulation of the biomolecular self-assembly.

Optimum excitation wavelength and photon energy threshold for spintronic terahertz emission from Fe/Pt bilayer.

Terahertz emission from ferromagnetic/non-magnetic spintronic heterostructures had been demonstrated as pump wavelength-independent. We report, however, the pump wavelength dependence of terahertz emission from an optimized Fe/Pt spintronic bilayer on MgO substrate. Maximum terahertz generation per total pump power was observed in the 1200- to 1800-nm pump wavelength range, and a marked decrease in the terahertz emission efficiency beyond 2500 nm (pump photon energies <0.5 eV) suggests a ∼0.35-eV threshold pump photon energy for effective spintronic terahertz emission. The inferred threshold is supported by previous theoretical results on the onset energy of significant spin-filtering at the Fe-Pt interface, and confirmed by Fe/Pt electronic structure calculations in this present work. The results of terahertz time-domain emission spectroscopy show the sensitivity of spintronic terahertz emission to both the optical absorptance of the heterostructure and the energy-dependent spin transport, as dictated by the properties of the metallic thin films.

Efficient electro-optic sampling detection of terahertz radiation via Cherenkov phase matching.

We experimentally demonstrate an efficient electro-optic sampling scheme based on Cherenkov phase matching of broadband terahertz radiation with 800-nm femtosecond probe beam in a 0.5 mm-thick LiNbO3 crystal coupled to a Si prism. The electro-optic signal from a Cherenkov-phase-matched LiNbO3 crystal is found to be comparable to that with a 4 mm-thick ZnTe crystal using a collinear phase matching. The Cherenkov phase matching technique can be achieved with any probe wavelength and hence has an advantage over the collinear phase matching method.

Machine learning for pattern and waveform recognitions in terahertz image data.

Several machine learning (ML) techniques were tested for the feasibility of performing automated pattern and waveform recognitions of terahertz time-domain spectroscopy datasets. Out of all the ML techniques under test, it was observed that random forest statistical algorithm works well with the THz datasets in both the frequency and time domains. With such ML algorithm, a classifier can be created with less than 1% out-of-bag error for segmentation of rusted and non-rusted sample regions of the image datasets in frequency domain. The degree of linear correlation between the rusted area percentage and the image spatial resolution with terahertz frequency can be used as an additional cross-validation criteria for the evaluation of classifier quality. However, for different rust staging measured datasets, a standardized procedure of image pre-processing is necessary to create/apply a single classifier and its usage is only limited to 1 ± 0.2 THz. Moreover, random forest is practically the best choice among the several popular ML techniques under test for waveform recognition of time-domain data in terms of classification accuracy and timing. Our results demonstrate the usefulness of random forest and several other machine learning algorithms for terahertz hyperspectral pattern recognition.

Dielectric property measurements of corneal tissues for computational dosimetry of the eye in terahertz band in vivo and in vitro.

The dielectric constant of the normal corneal tissue of a rabbit eye was obtained in vitro in the range from approximately 0.1 to 1 THz, and the drying process on the eye surface exposed to high-power terahertz waves was investigated by in vivo reflectance measurement using terahertz time-domain spectroscopy. When the rabbit eye was exposed to terahertz waves at 162 GHz for 6 min with an irradiation power of 360 or 480 mW/cm2, the reflectance temporally increased and then decreased with a temperature increase. Based on multiple-reflection calculation using the dielectric constant and anterior segment optical coherence tomography images, those changes in reflectance were attributed to drying of the tear and epithelium of the cornea, respectively. Furthermore, the drying progressed over a temperature increase of around 5°C under our exposure conditions. These findings suggest that the possibility of eye damage increases with the progress of drying and that the setting of the eye surface conditions can be a cause of disagreement between computational and experimental data of absorbed energy under high-level irradiation because reflectance is related to terahertz wave penetration in the eye tissue. The time-domain spectroscopic measurements were useful for the acquisition of the dielectric constant as well as for the real-time monitoring of the eye conditions during exposure measurement.