Transgenic sugarcane with higher levels of BRK1 showed improved drought tolerance.

KEY MESSAGE: Transgenic sugarcane overexpressing BRK1 showed improved tolerance to drought stress through modulation of actin polymerization and formation of interlocking marginal lobes in epidermal leaf cells, a typical feature associated with BRK1 expression under drought stress. BRICK1 (BRK1) genes promote leaf epidermal cell morphogenesis and division in plants that involves local actin polymerization. Although the changes in actin filament organization during drought have been reported, the role of BRK in stress tolerance remains unknown. In our previous work, the drought-tolerant Erianthus arundinaceus exhibited high levels of the BRK gene expression under drought stress. Therefore, in the present study, the drought-responsive gene, BRK1 from Saccharum spontaneum, was transformed into sugarcane to test if it conferred drought tolerance in the commercial sugarcane cultivar Co 86032. The transgenic lines were subjected to drought stress, and analyzed using physiological parameters for drought stress. The drought-induced BRK1-overexpressing lines of sugarcane exhibited significantly higher transgene expression compared with the wild-type control and also showed improved physiological parameters. In addition, the formation of interlocking marginal lobes in the epidermal leaf cells, a typical feature associated with BRK1 expression, was observed in all transgenic BRK1 lines during drought stress. This is the first report to suggest that BRK1 plays a role in sugarcane acclimation to drought stress and may prove to be a potential candidate in genetic engineering of plants for enhanced biomass production under drought stress conditions.

Overexpression of EaDREB2 and pyramiding of EaDREB2 with the pea DNA helicase gene (PDH45) enhance drought and salinity tolerance in sugarcane (Saccharum spp. hybrid).

KEY MESSAGE: EaDREB2 overexpressed in sugarcane enhanced tolerance to drought and salinity. When co-transformed with plant DNA helicase gene, DREB2 showed greater level of salinity tolerance than in single-gene transgenics. Drought is one of the most challenging agricultural issues limiting sustainable sugarcane production and can potentially cause up to 50 % yield loss. DREB proteins play a vital regulatory role in abiotic stress tolerance in plants. We previously reported that expression of EaDREB2 is enhanced by drought stress in Erianthus arundinaceus. In this study, we have isolated the DREB2 gene from E. arundinaceus, transformed one of the most popular sugarcane variety Co 86032 in tropical India with EaDREB2 through Agrobacterium-mediated transformation, pyramided the EaDREB2 gene with the gene coding for PDH45 driven by Port Ubi 2.3 promoter through particle bombardment and evaluated the V1 transgenics for soil deficit moisture and salinity stresses. Soil moisture stress was imposed at the tillering phase by withholding irrigation. Physiological, molecular and morphological parameters were used to assess drought tolerance. Salinity tolerance was assessed through leaf disc senescence and bud sprout assays under salinity stress. Our results indicate that overexpression of EaDREB2 in sugarcane enhances drought and salinity tolerance to a greater extent than the untransformed control plants. This is the first report of the co-transformation of EaDREB2 and PDH45 which shows higher salinity tolerance but lower drought tolerance than EaDREB2 alone. The present study seems to suggest that, for combining drought and salinity tolerance together, co-transformation is a better approach.

Interface magnetism in epitaxial BiFeO3-La0.7Sr0.3MnO3 heterostructures integrated on Si(100).

We report on the heteroepitaxial growth of ferroelectric (FE)-antiferromagnetic (AFM) BiFeO3 (BFO) on ferromagnetic La0.7Sr0.3MnO3 (LSMO), integrated on Si(100) using pulsed laser deposition via the domain matching epitaxy paradigm. The BFO/LSMO films were epitaxially grown on Si(100) by introducing epitaxial layers of SrTiO3/MgO/TiN. X-ray diffraction, scanning electron microscopy, high-resolution transmission electron microscopy, X-ray photo absorption spectroscopy, and atomic force microscopy were employed to fully characterize the samples. Furthermore, we have investigated the magnetic behavior of this five layer heterostructure, in which a d(5) system (Fe(3+)) manifested in FE-AFM BFO is epitaxially conjoined at the interface to a multivalent transition metal ion such as Mn(3+)/Mn(4+) in LSMO. The temperature- and magnetic field-dependent magnetization measurements reveal an unexpected enhancement in magnetic moment and improved magnetic hysteresis squareness originating from the BFO/LSMO interface. We observe a stronger temperature dependence of HEB when the polarity of field cooling is negative as compared to positive field cooling. We believe such an enhancement in magnetic moment and magnetic coupling is likely directly related to an electronic orbital reconstruction at the interface and complex interplay between orbital and spin degrees of freedom, similar to what has previously been reported in the literature. Future work will involve the linearly polarized X-ray absorption measurements to prove this hypothesis. This work represents a starting step toward the realization of magneto-electronic devices integrated with Si(100).

Serotype and genotype diversity of dengue viruses circulating in India: a multi-centre retrospective study involving the Virus Research Diagnostic Laboratory Network in 2018.

OBJECTIVES: A retrospective study was undertaken to investigate the circulating dengue virus (DENV) serotypes and genotypes in India in 2018. METHODS: In total, 4963 samples referred to virus research diagnostic laboratories (n=21), the Indian Council of Medical Research-National Institute of Virology (ICMR-NIV) and ICMR-NIV field units (n=2) for diagnosis of dengue in 2018 were tested using a real-time reverse transcription polymerase chain reaction assay for the presence of DENV serotypes. Representative samples were sequenced for the envelope (E) gene. RESULTS: Regional diversity was observed with regard to the dominant circulating serotypes. DENV-2 was found to be the most common serotype in many states. Thrombocytopenia, petechiae and malaise were associated with DENV-2 infection. Phylogenetic analyses of DENV E gene sequences revealed the circulation of genotypes I and V of DENV-1, two lineages of DENV-2 genotype IV, DENV-3 genotype III and DENV-4 genotype I. CONCLUSIONS: This study found regional differences in the prevalence of circulating DENV serotypes in India, and provides baseline data for continuous molecular surveillance. Molecular surveillance may have implications for predicting large-scale outbreaks of dengue if regional shifts in the predominantly circulating serotypes and genotypes are detected during the early phase of the dengue season.

Laser annealing of ion-implanted semiconductors.

The physical and electrical properties of ion-implanted silicon annealed with high-powered laser radiation are described. Particular emphasis is placed on the comparison of materials properties that can be achieved with laser annealing to those which can be achieved by conventional thermal annealing. Applications of these techniques to the fabrication of high-efficiency solar cells, and potential applications of this new technique to other materials areas are discussed.

Laser method for synthesis and processing of continuous diamond films on nondiamond substrates.

A laser method based upon carbon ion implantation and pulsed laser melting of copper has been used to produce continuous diamond thin film. Carbon ions were implanted with ion energies in the range of 60 to 120 keV, and doses of 1.0 x 10(18) to 2.0 x 10(18) ions cm(-2). The ion-implanted specimens were treated with nanosecond excimer laser pulses with the following parameters: energy density, 3.0 to 5.0 J cm(-2); wavelength, 0.308 microm; pulse width, 45 nanoseconds. The specimens were characterized with scanning electron microscopy (SEM), x-ray diffraction, Rutherford backscattering/ion channeling, Auger, and Raman spectroscopy. The macroscopic Raman spectra contained a strong peak at 1332 cm(-1) with full width at half maximum of 5 cm(-1), which is very close to the quality of the spectra obtained from single-crystal diamond. The selected area electron diffraction patterns and imaging confirmed the films to be defect-free single crystal over large areas of up to several square micrometers with no grain boundaries. Low voltage SEM imaging of surface features indicated the film to be continuous with presence of growth steps.

Magnetic properties of self-assembled Ni nanoparticles in two dimensional structures.

A pulsed laser deposition technique has been used to synthesize a uniform distribution of Ni nanoparticles of controllable size in Al2O3 thin film matrix. The ability to control particle size in confined layers provides a very convenient means to tune the magnetic properties from superparamagnetic to ferromagnetic. The coercivity of these particles was measured at various temperatures as a function of particle size. The results indicate that the magnetic transition from single- to multi-domain region occurs at a larger particle size at higher temperature than at lower temperature. Stronger magnetic interaction among particles at lower temperatures is believed to lead to the formation of smaller sized domains for any given particle size in order to minimize the interaction energy.

Direct conversion of carbon nanofibers and nanotubes into diamond nanofibers and the subsequent growth of large-sized diamonds.

We report a pulsed laser annealing method to convert carbon fibers and nanotubes into diamond fibers at ambient temperature and pressure in air. The conversion of carbon nanofibers and nanotubes into diamond nanofibers involves melting in a super undercooled state using nanosecond laser pulses, and quenching rapidly to convert into phase-pure diamond. The conversion process occurs at ambient temperature and pressure, and can be carried out in air. The structure of diamond fibers has been confirmed by selected-area electron diffraction in transmission electron microscopy, electron-back-scatter-diffraction in high-resolution scanning electron microscopy, all showing characteristic diffraction lines for the diamond structure. The bonding characteristics were determined by Raman spectroscopy with a strong peak near 1332 cm-1, and high-resolution electron-energy-loss spectroscopy in transmission electron microscopy with a characteristic peak at 292 eV for σ* for sp3 bonding and the absence of π* for sp2 bonding. The Raman peak at 1332 cm-1 downshifts to 1321 cm-1 for diamond nanofibers due to the phonon confinement in nanodiamonds. These laser-treated carbon fibers with diamond seeds are used to grow larger diamond crystallites further by using standard hot-filament chemical vapor deposition (HFCVD). We compare these results with those obtained without laser treating the carbon fibers. The details of diamond conversion and HFCVD growth are presented in this paper.

In silico characterisation and functional validation of chilling tolerant divergence 1 (COLD1) gene in monocots during abiotic stress.

The G protein-coupled receptor is one of the major transmembrane proteins in plants. It consists of an α subunit, a ß subunit and three γ subunits. Chilling tolerant divergence 1 (COLD1) includes a Golgi pH receptor (GPHR) domain, which maintains cell membrane organisation and dynamics, along with abscisic acid linked G protein-coupled receptor (ABA_GPCR) that regulates the signalling pathways during cold stress. In the present study, we performed characterisation of a homologous COLD1 from the economically important monocot species Oryza sativa L., Zea mays L., Sorghum bicolor (L.)Moench and Erianthus arundinaceus (L.) Beauv. IK 76-81, a wild relative of Saccharum. COLD1 was isolated from E. arundinaceus IK 76-81, analysed for its evolution, domain, membrane topology, followed by prediction of secondary, tertiary structures and functionally validated in all four different monocots. Gene expression studies of COLD1 revealed differential expression under heat, drought, salinity and cold stresses in selected monocots. This is the first study on regulation of native COLD1 during abiotic stress in monocots, which has opened up new leads for trait improvement strategies in this economically important crop species.

Corrigendum to: In silico characterisation and functional validation of chilling tolerant divergence 1 (COLD1) gene in monocots during abiotic stress.

The G protein-coupled receptor is one of the major transmembrane proteins in plants. It consists of an α subunit, a ß subunit and three γ subunits. Chilling tolerant divergence 1 (COLD1) includes a Golgi pH receptor (GPHR) domain, which maintains cell membrane organisation and dynamics, along with abscisic acid linked G protein-coupled receptor (ABA_GPCR) that regulates the signalling pathways during cold stress. In the present study, we performed characterisation of a homologous COLD1 from the economically important monocot species Oryza sativa L., Zea mays L., Sorghum bicolor (L.)Moench and Erianthus arundinaceus (L.) Beauv. IK 76-81, a wild relative of Saccharum. COLD1 was isolated from E. arundinaceus IK 76-81, analysed for its evolution, domain, membrane topology, followed by prediction of secondary, tertiary structures and functionally validated in all four different monocots. Gene expression studies of COLD1 revealed differential expression under heat, drought, salinity and cold stresses in selected monocots. This is the first study on regulation of native COLD1 during abiotic stress in monocots, which has opened up new leads for trait improvement strategies in this economically important crop species.

Isolation and characterization of water-deficit stress-responsive α-expansin 1 (EXPA1) gene from Saccharum complex.

In this study, full-length (1282-1330 bp) α-expansin 1 (EXPA1) gene from three different accessions belonging to Saccharum complex (Saccharum officinarum-SoEXPA1, Erianthus arundinaceus-EaEXPA1, and Saccharum spp. hybrid-ShEXPA1) was isolated using RAGE technique and characterized. The intronic and coding regions of isolated expansin genes ranged between 526-568 and 756-762 bp, respectively. An open reading frame encoding a polypeptide of 252 amino acids was obtained from S. officinarum and commercial sugarcane hybrid, whereas 254 amino acids were obtained in E. arundinaceus, a wild relative of Saccharum. Bioinformatics analysis of deduced protein revealed the presence of specific signature sequences and conserved amino acid residues crucial for the functioning of the protein. The predicted physicochemical characterization showed that the protein is stable in nature with instability index (II) value less than 40 and also clearly shown the dominance of random coil in the protein structure. Phylogenetic analysis revealed high conservation of EXPA1 among Saccharum complex and related crop species, Sorghum bicolor and Zea mays. The docking study of EXPA1 protein showed the interaction with xylose, which is present in xyloglucan of plant cell wall, elucidated the role of the expansin proteins in plant cell wall modification. This was further supported by the subcellular localization experiment in which it is clearly seen that the expansin protein localizes in the cell wall. Relative expression analysis of EXPA1 gene in Saccharum complex during drought stress showed high expression of the EaEXPA1 in comparison with SoEXPA1 and ShEXPA1 indicating possible role of EaEXPA1 in increased water-deficit stress tolerance in E. arundinaceus. These results suggest the potential use of EXPA1 for increasing the water-deficient stress tolerance levels in crop plants.

Comparative Raman and HRTEM study of nanostructured GaN nucleation layers and device layers on sapphire (0001).

Raman spectroscopy in conjunction with high-resolution transmission electron microscopy (HRTEM) has been used to study structural characteristics and strain distribution of the nanostructured GaN nucleation layer (NL) and the GaN device layer on (0001) sapphire substrates used for light-emitting diodes and lasers. Raman peaks corresponding to the cubic and the hexagonal phase of GaN are observed in the Raman spectrum from 15 nm and 45 nm NLs. A comparison of the peak intensities for the cubic and hexagonal phases of GaN in the NLs suggests that the cubic phase is dominant in the 15 nm NL and the hexagonal phase in the 45 nm NL. An increase in the density of stacking faults in the metastable cubic GaN (c-GaN) phase with increasing growth time lowers the system energy as well as locally converts c-GaN phase into hexagonal GaN (h-GaN). It also explains the observation of the more intense peaks of h-GaN in the 45 nm NL compared to c-GaN peaks. For the sample wherein an h-GaN device layer was grown at higher temperatures on the NL, narrow Raman peaks corresponding to only h-GaN were observed, confirming the high-quality of the films. The peak shift of the E2(H)(LO) mode of h-GaN in the NLs and the h-GaN film suggests the presence of a tensile stress in the NL which is attributed to defects such as stacking faults and twins, and a compressive stress in high-temperature grown h-GaN film which is attributed to the thermal-expansion mismatch between the film and the substrate. The peak shifts of the substrate also reveal that during the low temperature growth of the NL the substrate is under a compressive stress which is attributed to defects in the NL and during the high temperature growth of the device layer, there is a tensile strain in the substrate as expected from differences in coefficients of thermal expansion of the film and the substrate during the cooling cycle.

Nanostructured GaN nucleation layer for light-emitting diodes.

This paper addresses the formation of nanostructured gallium nitride nucleation (NL) or initial layer (IL), which is necessary to obtain a smooth surface morphology and reduce defects in h-GaN layers for light-emitting diodes and lasers. From detailed X-ray and HR-TEM studies, researchers determined that this layer consists of nanostructured grains with average grain size of 25 nm, which are separated by small-angle grain boundaries (with misorientation approximately 1 degrees), known as subgrain boundaries. Thus NL is considered to be single-crystal layer with mosaicity of about 1 degrees. These nc grains are mostly faulted cubic GaN (c-GaN) and a small fraction of unfaulted c-GaN. This unfaulted Zinc-blende c-GaN, which is considered a nonequilibrium phase, often appears as embedded or occluded within the faulted c-GaN. The NL layer contained in-plane tensile strain, presumably arising from defects due to island coalescence during Volmer-Weber growth. The 10L X-ray scans showed c-GaN fraction to be over 63% and the rest h-GaN. The NL layer grows epitaxially with the (0001) sapphire substrate by domain matching epitaxy, and this epitaxial relationship is remarkably maintained when c-GaN converts into h-GaN during high-temperature growth.

Structural, magnetic, and electron transport studies on nanocrystalline layered manganite La1.2Ba1,8Mn2O7 system.

The structure property relationship for Ru doped La1.2Ba1.8Mn2-RuxO7 system has been studied systematically. The system crystallizes in the single-phase tetragonal structure with space group of 14/mmm. The unit cell volume is found to increase with Ru doping. The sheet type microstructure could be seen in this system, which is important for anisotropic nature of layered structure. The crystallite size is found to be 25 nm indicating nanocrystalline nature of the system. Ferromagnetic to paramagnetic (Tc) transition above the room temperature is observed in all except the highest doped Ru (x = 1.0) where the Tc is 254 K using a.c. susceptibility measurement. The large values of magnetoresistance for the x = 0.0 sample at 10 K is found to be 57% and 64% at applied fields of 5 and 10 T, respectively.

Growth and observation of low-field giant magnetoresistance in La0.7Sr0.3MnO3/ZnO superlattice structures.

We report the growth of a new class of superlattice structure, consisting of alternate layers of La0.7Sr0.3MnO3 (LSMO) and ZnO, which exhibits giant magnetoresistance at low fields. These superlattices were fabricated using a novel pulsed-laser deposition technique with a specially designed target assembly. Giant magnetoresistance of > 250% has been observed in these structures in current-in-plane configuration on the application of just -400 Gauss of magnetic field over the broad temperature range 15-200 K. Observation of giant magnetoresistance at such low magnetic fields is a groundbreaking step in the field of novel magnetic materials and devices.

Introduction of Pea DNA Helicase 45 Into Sugarcane (Saccharum spp. Hybrid) Enhances Cell Membrane Thermostability And Upregulation Of Stress-responsive Genes Leads To Abiotic Stress Tolerance.

DNA helicases are motor proteins that play an essential role in nucleic acid metabolism, by providing a duplex-unwinding function. To improve the drought and salinity tolerance of sugarcane, a DEAD-box helicase gene isolated from pea with a constitutive promoter, Port Ubi 2.3 was transformed into the commercial sugarcane variety Co 86032 through Agrobacterium-mediated transformation, and the transgenics were screened for tolerance to soil moisture stress and salinity. The transgene integration was confirmed through polymerase chain reaction, and the V 0 transgenic events showed significantly higher cell membrane thermostability under normal irrigated conditions. The V 1 transgenic events were screened for tolerance to soil moisture stress and exhibited significantly higher cell membrane thermostability, transgene expression, relative water content, gas exchange parameters, chlorophyll content, and photosynthetic efficiency under soil moisture stress compared to wild-type (WT). The overexpression of PDH45 transgenic sugarcane also led to the upregulation of DREB2-induced downstream stress-related genes. The transgenic events demonstrated higher germination ability and better chlorophyll retention than WT under salinity stress. Our results suggest the possibility for development of increased abiotic stress tolerant sugarcane cultivars through overexpression of PDH45 gene. Perhaps this is the first report, which provides evidence for increased drought and salinity tolerance in sugarcane through overexpression of PDH45.

Erianthus arundinaceus HSP70 (EaHSP70) overexpression increases drought and salinity tolerance in sugarcane (Saccharum spp. hybrid).

Heat shock proteins (HSPs) have a major role in stress tolerance mechanisms in plants. Our studies have shown that the expression of HSP70 is enhanced under water stress in Erianthus arundinaceus. In this paper, we evaluate the effects of overexpression of EaHSP70 driven by Port Ubi 2.3 promoter in sugarcane. The transgenic events exhibit significantly higher gene expression, cell membrane thermostability, relative water content, gas exchange parameters, chlorophyll content and photosynthetic efficiency. The overexpression of EaHSP70 transgenic sugarcane led to the upregulation of stress-related genes. The transformed sugarcane plants had better chlorophyll retention and higher germination ability than control plants under salinity stress. Our results suggest that EaHSP70 plays an important role in sugarcane acclimation to drought and salinity stresses and its potential for genetic engineering of sugarcane for drought and salt tolerance.

Acute interstitial nephritis and acute renal failure associated with cimetidine therapy.

We are reporting the clinical and pathologic features of severe acute renal failure which developed in a patient after the institution of cimetidine (Targamet) therapy. A renal biopsy specimen revealed an acute interstitial nephritis with numerous plasma cells and eosinophils suggestive of an immunologic reaction. Marked improvement of renal function occurred within three weeks after cessation of the drug therapy. Our clinical and pathologic data strongly suggest an association between acute interstitial nephritis and cimetidine, a suggestion which should be confirmed by additional case studies.

Novel methods of forming self-assembled nanostructured materials: Ni nanodots in Al2O3 and TiN matrices.

We have developed two novel methods of growing self-assembled nanodot arrays of uniform size (diameter 2-50 nm) of a variety of materials in crystalline as well as amorphous matrices. These methods employ a pulsed laser deposition technique and are classified as: (a) sequential growth method; and (b) simultaneous growth method. In the first method, the nanodots and matrix material are formed sequentially, while in the second method, nanodots and matrix materials grow simultaneously. In the sequential growth, self-assembly of nanodots is controlled by the flux of materials, interfacial energy, intervening matrix layer, substrate and laser parameters. For the simultaneous growth method of self-assembly of nanodots, there is additional requirement that the material of the nanodot and the matrix should be such that the Gibb's free energy of oxidation of the constituent of matrix material is much lower than that of the nanodot material.

Role of self-assembled gold nanodots in improving the electrical and optical characteristics of zinc oxide films.

We have studied the effect of embedding nanocrystalline Au particles on the electrical and optical characteristics of ZnO films. Au-embedded epitaxial ZnO films were deposited on (0001) sapphire substrates with a pulsed laser deposition technique. The crystalline quality of both the ZnO matrix and Au nanoparticles was investigated by X-ray diffraction and transmission electron microscopy. Composite films were characterized by photoluminescence, optical absorption, and low-temperature electrical resistivity measurements. Photoluminescence spectra of theses films showed a sharp excitonic peak at 3.22 +/- 0.05 eV without any signature of green band emission. Electrical resistivity measurements showed these films to be highly conducting, with a room-temperature resistivity of 3.4 +/- 0.2 m omega-cm.