Catalase associated with antagonistic changes of abscisic acid and gibberellin response, biosynthesis and catabolism is involved in eugenol-inhibited seed germination in rice.

KEY MESSAGE: The inhibitory effect of eugenol on rice germination is mediated by a two-step modulatory process: Eugenol first regulates the antagonism of GA and ABA, followed by activation of catalase activity. The natural monoterpene eugenol has been reported to inhibit preharvest sprouting in rice. However, the inhibitory mechanism remains obscure. In this study, simultaneous monitoring of GA and ABA responses by the in vivo GA and ABA-responsive dual-luciferase reporter system showed that eugenol strongly inhibited the GA response after 6 h of imbibition, whereas eugenol significantly enhanced the ABA response after 12 h of imbibition. Gene expression analysis revealed that eugenol significantly induced the ABA biosynthetic genes OsNCED2, OsNCED3, and OsNCED5, but notably suppressed the ABA catabolic genes OsABA8ox1 and OsABA8ox2. Conversely, eugenol inhibited the GA biosynthetic genes OsGA3ox2 and OsGA20ox4 but significantly induced the GA catabolic genes OsGA2ox1 and OsGA2ox3 during imbibition. OsABI4, the key signaling regulator of ABA and GA antagonism, was notably induced before 12 h and suppressed after 24 h by eugenol. Moreover, eugenol markedly reduced the accumulation of H2O2 in seeds after 36 h of imbibition. Further analysis showed that eugenol strongly induced catalase activity, protein accumulation, and the expression of three catalase genes. Most importantly, mitigation of eugenol-inhibited seed germination was found in the catc mutant. These findings indicate that catalase associated with antagonistic changes of ABA and GA is involved in the sequential regulation of eugenol-inhibited seed germination in rice.

On-Chip Monolithically Integrated Ultraviolet Low-Threshold Plasmonic Metal-Semiconductor Heterojunction Nanolasers.

The metal-semiconductor heterojunction is imperative for the realization of electrically driven nanolasers for chip-level platforms. Progress in developing such nanolasers has hitherto rarely been realized, however, because of their complexity in heterojunction fabrication and the need to use noble metals that are incompatible with microelectronic manufacturing. Most plasmonic nanolasers lase either above a high threshold (101 -103 MW cm-2 ) or at a cryogenic temperature, and lasing is possible only after they are removed from the substrate to avoid the large ohmic loss and the low modal reflectivity, making monolithic fabrication impossible. Here, for the first time, record-low-threshold, room-temperature ultraviolet (UV) lasing of plasmon-coupled core-shell nanowires that are directly grown on silicon is demonstrated. The naturally formed core-shell metal-semiconductor heterostructure of the nanowires leads to a 100-fold improvement in growth density over previous results. This unprecedentedly high nanowire density creates intense plasmonic resonance, which is outcoupled to the resonant Fabry-Pérot microcavity. By boosting the emission strength by a factor of 100, the hybrid photonic-plasmonic system successfully facilitates a record-low laser threshold of 12 kW cm-2 with a spontaneous emission coupling factor as high as ≈0.32 in the 340-360 nm range. Such architecture is simple and cost-competitive for future UV sources in silicon integration.

The Influence of Helium Dielectric Barrier Discharge Jet (DBDjet) Plasma Treatment on Bathocuproine (BCP) in p-i-n-Structure Perovskite Solar Cells.

A bathocuproine (BCP) layer is typically used as the hole-blocking layer in p-i-n-structure perovskite solar cells (PSCs) between PC61BM and Ag electrodes. Before evaporating the Ag, we used a low-temperature (<40 °C) atmospheric-pressure dielectric barrier discharge jet (DBDjet) to treat the BCP with different scan rates. The main purpose of this was to change the contact resistance between the BCP layer and the Ag electrodes through surface modification using a DBDjet. The best power conversion efficiency (PCE) of 13.11% was achieved at a DBDjet scan rate of 2 cm/s. The He DBDjet treatment introduced nitrogen to form C-N bonds and create pits on the BCP layer. This deteriorated the interface between the BCP and the follow-up deposited-Ag top electrode. Compared to the device without the plasma treatment on the BCP layer, the He DBDjet treatment on the BCP layer reduced photocurrent hysteresis but deteriorated the fill factor and the efficiency of the PSCs.

Near-Infrared Emission Induced by Shortened Pt-Pt Contact: Diplatinum(II) Complexes with Pyridyl Pyrimidinato Cyclometalates.

Four diplatinum(II) complexes with the formula [Pt(pypm)(µ-Fn)]2 (2, 3a-c) bearing both a pyridine-pyrimidinate chelate and formamidinate bridge, where (pypm)H and FnH stand for 5-(pyridin-2-yl)-2-(trifluoromethyl)pyrimidine and functional formamidines with various substituents of iPr (n = 1), Ph (n = 2), C6H4tBu (n = 3), and C6H4CF3 (n = 4), were synthesized en route from a mononuclear intermediate represented by [Pt(pypm)Cl(F1H)] (1). Single-crystal X-ray diffraction studies confirmed the structure of 1 and 3a comprised of an individual "Pt(pypm)" unit and two "Pt(pypm)" units with a Pt···Pt distance of 2.8845(2) Å, respectively. Therefore, in contrast to the structured emission of mononuclear 1 with the first vibronic peak wavelength at 475 nm, all other diplatinum complexes with shortened Pt···Pt separation exhibited greatly red shifted and structureless metal-metal to ligand charge transfer (MMLCT) emission that extended into the near-infrared region in solid states. Their photophysical characteristics were measured under three distinctive morphological states (i.e., crystals, sublimed powders, and vacuum-deposited thin films) by steady-state UV-vis spectroscopy, while retention of Pt···Pt interactions in deposited thin films of 2 and 3a-c was confirmed using Raman spectroscopy, demonstrating lowered Pt···Pt stretching at 80-200 cm-1. Most importantly, complexes 3a-c exhibited a gradual red shift with the trends crystals < sublimed powders < vacuum-deposited thin films, a result of increased intermolecular π-π stacking interactions and Pt···Pt interactions, while crystalline samples exhibited the highest luminescence among all three morphological states due to the fewest defects in comparison to other morphologies. Finally, 3b was selected as a nondoped emitter for the fabrication of NIR-emitting OLEDs, giving an electroluminescence peak at 767 nm and a maximum external quantum efficiency of 0.14% with negligible roll-off.

H2O2-Based Method for Rapid Detection of Transgene-Free Rice Plants from Segregating CRISPR/Cas9 Genome-Edited Progenies.

Genome-editing techniques such as CRISPR/Cas9 have been widely used in crop functional genomics and improvement. To efficiently deliver the guide RNA and Cas9, most studies still rely on Agrobacterium-mediated transformation, which involves a selection marker gene. However, several limiting factors may impede the efficiency of screening transgene-free genome-edited plants, including the time needed to produce each life cycle, the response to selection reagents, and the labor costs of PCR-based genotyping. To overcome these disadvantages, we developed a simple and high-throughput method based on visual detection of antibiotics-derived H2O2 to verify transgene-free genome-edited plants. In transgenic rice containing hygromycin phosphotransferase (HPT), H2O2 content did not change in the presence of hygromycin B (HyB). In contrast, in transgenic-free rice plants with 10-h HyB treatment, levels of H2O2 and malondialdehyde, indicators of oxidative stress, were elevated. Detection of H2O2 by 3,3'-diaminobenzidine (DAB) staining suggested that H2O2 could be a marker to efficiently distinguish transgenic and non-transgenic plants. Analysis of 24 segregating progenies of an HPT-containing rice plant by RT-PCR and DAB staining verified that DAB staining is a feasible method for detecting transformants and non-transformants. Transgene-free genome-edited plants were faithfully validated by both PCR and the H2O2-based method. Moreover, HyB induced overproduction of H2O2 in leaves of Arabidopsis, maize, tobacco, and tomato, which suggests the potential application of the DAB method for detecting transgenic events containing HPT in a wide range of plant species. Thus, visual detection of DAB provides a simple, cheap, and reliable way to efficiently identify transgene-free genome-edited and HPT-containing transgenic rice.

Effects of light intensity and plant growth regulators on callus proliferation and shoot regeneration in the ornamental succulent Haworthia.

BACKGROUND: Haworthia are desert succulents belonging to the Asphodelaceae family. Haworthia species are cultivated commercially as ornamentals and some rare species are quite valuable at retail market but growth slowly and difficult to propagation. However, an efficient micropropagation protocol was remained insufficient. RESULTS: The organogenic cultures obtained from inflorescence explants were cultured on Murashige and Skoog (MS) medium supplemented with various combinations of 6-benzylaminopurine (BA) and α-naphthalene acetic acid (NAA) under a light intensity of 10 µmol m-2 s-1 or 45 µmol m-2 s-1. The highest callus proliferation index (93.15%) with 1.0 mg L-1 BA + 0.1 mg L-1 NAA under a light intensity of 10 µmol m-2 s-1. The best shoot proliferation rates were on media with either 1 mg L-1 BA + 0-0.4 mg L-1 NAA (65.57-81.01%) under a light intensity of 45 µmol m-2 s-1. The highest root length (15.57 mm) and the highest rooting frequency (17 roots per shoot) were obtained when adventitious shoots were inoculated on MS medium with 0.4 mg L-1 NAA + 0.4 mg L-1 IBA. The survival rate of the transplanted plantlets was about 100%. The efficient micropropagation protocol proliferated Haworthia regenerate plants from inflorescence within 11 weeks. CONCLUSIONS: The present study determined the best combination of light intensity and plant growth regulators (PGRs) for improved organogenesis of Haworthia during propagation by tissue culture. This optimized protocol showed light intensity is an important factor for efficient callus or shoot regeneration. These results indicate that it will be useful to optimize the light conditions for future commercial cultivation, germplasm conservation, genetic engineering and molecular biology research of this ornamental plant.

MIPs-graphene nanoplatelets-MWCNTs modified glassy carbon electrode for the determination of cardiac troponin I.

An electrochemical sensor with high selectivity in addition to sensitivity was developed for the determination of cardiac troponin I (cTnI), based on the modification of cTnI imprinted polymer film on a glassy carbon electrode (GCE). The sensor was fabricated by layer-by-layer assembled graphene nanoplatelets (GS), multiwalled carbon nanotubes (MWCNTs), chitosan (CS), glutaraldehyde (GA) composites, which can increase the electronic transfer rate and the active surface area to capture a larger number of antigenic proteins. MWCNTs/GS based imprinted polymers (MIPs/MWCNTs/GS) were synthesized by means of methacrylic acid (MAA) as the monomer, ethylene glycol dimethacrylate (EGDMA) as the cross linker α,α'-azobisisobutyronitrile (AIBN) as the initiator and cTnI as the template. In comparison with conventional methods, the proposed electrochemical sensor is highly sensitive for cTnI, providing a better linear response range from 0.005 to 60 ng cm-3 and a lower limit of detection (LOD) of 0.0008 ng cm-3 under optimal experimental conditions. In addition, the electrochemical sensor exhibited good specificity, acceptable reproducibility and stability. Moreover, satisfactory results were obtained in real human serum samples, indicating that the developed method has the potential to find application in clinical detection of cTnI as an alternative approach.

Atomically smooth hybrid crystalline-core glass-clad fibers for low-loss broadband wave guiding.

We demonstrate direct evidence for the first realization of atomically smooth sapphire crystalline fiber cores with a surface variation of only ~1.9 Å. The hybrid glass-clad crystalline cores were grown by a laser-based fiber drawing technique. Because of the improvement in crystal fiber quality, we were able, for the first time, to comprehensively and quantitatively elucidate the correlation between fiber nanostructure and optical loss. We also experimentally demonstrated that high-temperature treatment has a significant impact on defect relaxation and promotes excellent crystallinity, and hence enables low-loss optical wave guiding. The experimentally measured propagation losses in the order of 0.01-0.1 dB/cm are the lowest ever reported among conventional Ti:sapphire channel waveguides and ultrafast-laser-inscribed waveguides, and agree well with the theory. Through experiments and numerical calculation, we have demonstrated that low threshold and high efficiency of Ti:sapphire crystal fiber lasers are possible with the atomic-level roughness, low-loss propagation, and high crystallinity of the Ti:sapphire crystalline core.

The genome and transcriptome of Phalaenopsis yield insights into floral organ development and flowering regulation.

The Phalaenopsis orchid is an important potted flower of high economic value around the world. We report the 3.1 Gb draft genome assembly of an important winter flowering Phalaenopsis 'KHM190' cultivar. We generated 89.5 Gb RNA-seq and 113 million sRNA-seq reads to use these data to identify 41,153 protein-coding genes and 188 miRNA families. We also generated a draft genome for Phalaenopsis pulcherrima 'B8802,' a summer flowering species, via resequencing. Comparison of genome data between the two Phalaenopsis cultivars allowed the identification of 691,532 single-nucleotide polymorphisms. In this study, we reveal that the key role of PhAGL6b in the regulation of labellum organ development involves alternative splicing in the big lip mutant. Petal or sepal overexpressing PhAGL6b leads to the conversion into a lip-like structure. We also discovered that the gibberellin pathway that regulates the expression of flowering time genes during the reproductive phase change is induced by cool temperature. Our work thus depicted a valuable resource for the flowering control, flower architecture development, and breeding of the Phalaenopsis orchids.

A de novo floral transcriptome reveals clues into Phalaenopsis orchid flower development.

Phalaenopsis has a zygomorphic floral structure, including three outer tepals, two lateral inner tepals and a highly modified inner median tepal called labellum or lip; however, the regulation of its organ development remains unelucidated. We generated RNA-seq reads with the Illumina platform for floral organs of the Phalaenopsis wild-type and peloric mutant with a lip-like petal. A total of 43,552 contigs were obtained after de novo assembly. We used differentially expressed gene profiling to compare the transcriptional changes in floral organs for both the wild-type and peloric mutant. Pair-wise comparison of sepals, petals and labellum between peloric mutant and its wild-type revealed 1,838, 758 and 1,147 contigs, respectively, with significant differential expression. PhAGL6a (CUFF.17763), PhAGL6b (CUFF.17763.1), PhMADS1 (CUFF.36625.1), PhMADS4 (CUFF.25909) and PhMADS5 (CUFF.39479.1) were significantly upregulated in the lip-like petal of the peloric mutant. We used real-time PCR analysis of lip-like petals, lip-like sepals and the big lip of peloric mutants to confirm the five genes' expression patterns. PhAGL6a, PhAGL6b and PhMADS4 were strongly expressed in the labellum and significantly upregulated in lip-like petals and lip-like sepals of peloric-mutant flowers. In addition, PhAGL6b was significantly downregulated in the labellum of the big lip mutant, with no change in expression of PhAGL6a. We provide a comprehensive transcript profile and functional analysis of Phalaenopsis floral organs. PhAGL6a PhAGL6b, and PhMADS4 might play crucial roles in the development of the labellum in Phalaenopsis. Our study provides new insights into how the orchid labellum differs and why the petal or sepal converts to a labellum in Phalaenopsis floral mutants.

Molecular characterization of the Oncidium orchid HDR gene encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase, the last step of the methylerythritol phosphate pathway.

Two pathways are used by higher plants for the biosynthesis of isoprenoid precursors: the mevalonate pathway in the cytosol and a 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway in the plastids, with 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (HDR) catalyzing the last step in the MEP pathway. In order to understand the contribution of MEP pathway in isoprenoid biosynthesis of Oncidium orchid, a full-length cDNA corresponding to HDR from the flower tissues of Oncidium Gower Ramsey was cloned. The deduced OncHDR amino acid sequence contains a plastid signal peptide at the N-terminus and four conserved cysteine residues. RT-PCR analysis of HDR in Oncidium flowering plants revealed ubiquitous expression in organs and tissues, with preferential expression in the floral organs. Phylogenetic analysis revealed evolutionary conservation of the encoding HDR protein sequence. The genomic sequence of the HDR in Oncidium is similar to that in Arabidopsis, grape, and rice in structure. Successful complementation by OncHDR of an E. coli hdr(-) mutant confirmed its function. Transgenic tobacco carrying the OncHDR promoter-GUS gene fusion showed expression in most tissues, as well as in reproductive organs, as revealed by histochemical staining. Light induced strong GUS expression driven by the OncHDR promoter in transgenic tobacco seedlings. Taken together, our data suggest a role for OncHDR as a light-activated gene.

Transcription analysis of peloric mutants of Phalaenopsis orchids derived from tissue culture.

Tissue culture has been widely used for mass propagation of Phalaenopsis. However, somaclonal variation occurred during micropropagation process posed a severe problem by affecting product quality. In this study, wild type and peloric flower buds of Phalaenopsis hybrids derived from flower stalk nodal culture were used for cDNA-RAPD and cDNA suppression subtractive hybridization analyses in order to study their genetic difference in terms of expressed sequence tags. A total of 209 ESTs from normal flower buds and 230 from mutants were sequenced. These ESTs sequences can be grouped into several functional categories involved in different cellular processes including metabolism, signal transduction, transcription, cell growth and division, protein synthesis, and protein localization, and into a subcategory of proteins with unknown function. Cymbidium mosaic virus transcript was surprisingly found expressed frequently in the peloric mutant of P. Little Mary. Real-time RT-PCR analysis on selected ESTs showed that in mutant flower buds, a bZIP transcription factor (TGA1a-like protein) was down-regulated, while up-regulated genes include auxin-regulated protein kinase, cyclophilin, and TCP-like genes. A retroelement clone was also preferentially expressed in the peloric mutant flowers. On the other hand, ESTs involved in DNA methylation, chromatin remodeling and post-transcriptional regulation, such as DNA methyltransferase, histone acetyltransferase, ERECTA, and DEAD/DEAH RNA helicase, were enriched in normal flower buds than the mutants. The enriched transcripts in the wild type indicate the down regulation of these transcripts in the mutants, and vice versa. The potential roles of the analyzed transcripts in the development of Phalaenopsis flowers are discussed.