An extremely bad-cavity laser

Lasing in the bad-cavity regime has promising applications in precision measurement and frequency metrology due to the reduced sensitivity of the laser frequency to cavity length fluctuations. Thus far, relevant studies have been mainly focused on conventional cavities whose finesse is high enough that the resonance linewidth is sufficiently narrow compared to the cavity's free spectral range, though still in the bad-cavity regime. However, lasing output from the cavity whose finesse is close to the limit of 2 has never been experimentally accessed. Here, we demonstrate an extremely bad-cavity laser, analyze the physical mechanisms limiting cavity finesse, and report on the worst ever laser cavity with finesse reaching 2.01. The optical cavity has a reflectance close to zero and only provides a weak optical feedback. The laser power can be as high as tens of $\mu$W and the spectral linewidth reaches a few kHz, over one thousand times narrower than the gain bandwidth. In addition, the measurement of cavity pulling reveals a pulling coefficient of 0.0148, the lowest value ever achieved for a continuous wave laser. Our findings open up an unprecedentedly innovative perspective for future new ultra-stable lasers, which could possibly trigger the future discoveries in optical clocks, cavity QED, continuous wave superradiant laser, and explorations of quantum manybody physics

Overexpression of the Glutathione Peroxidase 5 (RcGPX5) Gene From Rhodiola crenulata Increases Drought Tolerance in Salvia miltiorrhiza

Excessive cellular accumulation of reactive oxygen species (ROS) due to environmental stresses can critically disrupt plant development and negatively affect productivity. Plant glutathione peroxidases (GPXs) play an important role in ROS scavenging by catalyzing the reduction of H2O2 and other organic hydroperoxides to protect plant cells from oxidative stress damage. RcGPX5, a member of the GPX gene family, was isolated from a traditional medicinal plant Rhodiola crenulata and constitutively expressed in Salvia miltiorrhiza under control of the CaMV 35S promoter. Transgenic plants showed increased tolerance to oxidative stress caused by application of H2O2 and drought, and had reduced production of malondialdehyde (MDA) compared with the wild type. Under drought stress, seedlings of the transgenic lines wilted later than the wild type and recovered growth 1 day after re-watering. In addition, the reduced glutathione (GSH) and total glutathione (T-GSH) contents were higher in the transgenic lines, with increased enzyme activities including glutathione reductase (GR), ascorbate peroxidase (APX), and GPX. These changes prevent H2O2 and O2- accumulation in cells of the transgenic lines compared with wild type. Overexpression of RcGPX5 alters the relative expression levels of multiple endogenous genes in S. miltiorrhiza, including transcription factor genes and genes in the ROS and ABA pathways. In particular, RcGPX5 expression increases the mass of S. miltiorrhiza roots while reducing the concentration of the active ingredients. These results show that heterologous expression of RcGPX5 in S. miltiorrhiza can affect the regulation of multiple biochemical pathways to confer tolerance to drought stress, and RcGPX5 might act as a competitor with secondary metabolites in the S. miltiorrhiza response to environmental stimuli

Genetic Mapping of Prince Rupprecht’s Larch (Larix principis-rupprechtii Mayr) by Specific-Locus Amplified Fragment Sequencing

A high-density genetic linkage map is essential for plant genetics and genomics research. However, due to the deficiency of genomic data and high-quality molecular markers, no genetic map has been published for Prince Rupprecht’s larch (Larix principis-rupprechtii Mayr), a conifer species with high ecological and commercial value in northern China. In this study, 145 F1 progeny individuals from an intraspecific cross between two elite clones of L. principis-rupprechtii and their parents were employed to construct the first genetic map in this important tree species using specific-locus amplified fragment sequencing (SLAF-seq). After preprocessing, the procedure yielded 300.20 Gb of raw data containing 1501.22 M pair-end reads. A total of 324,352 SNP markers were detected and 122,785 of them were polymorphic, with a polymorphism rate of 37.86%. Ultimately, 6099 SNPs were organized into a genetic map containing 12 linkage groups, consistent with the haploid chromosome number of larch and most other species in the Pinaceae family. The linkage map spanned 2415.58 cM and covered 99.6% of the L. principis-rupprechtii genome with an average of 0.4 cM between adjacent markers. To the best of our knowledge, this map is the first reference map for L. principis-rupprechtii, as well as the densest one obtained in larch species thus far. The genome-wide SNPs and the high-resolution genetic map will provide a foundation for future quantitative trait loci mapping, map-based cloning, marker-assisted selection, comparative genomics, and genome sequence assembly for larch trees

MrSVP, a secreted virulence-associated protein, contributes to thermotolerance and virulence of the entomopathogenic fungus Metarhizium robertsii

Abstract Background Metarhizium robertsii, a widely distributed insect pathogen, is presently used as a natural alternative to chemical insecticides. Unfortunately, its worldwide commercial use has been restricted by a short shelf life and inconsistencies in virulence. In our previous study, a gene (GenBank accession number EFZ01626) was found to be significantly upregulated in heat-treated conidia. In the present study, this gene was characterized via gene disruption and complementation strategies. Results The gene (amplified by rapid amplification of cDNA ends PCR) was 1219 bp long and contained an open reading frame (ORF) of 777 bp. It encoded a protein of 234 amino acid residues with a 26-residue signal peptide. Bioinformatics analyses did not identify conserved functional domains; therefore, it was assumed to be a secreted virulence-associated protein according to its signal peptide and bioassay results. We found that the conidial germination rate of the ΔMrSVP mutant fungi dramatically decreased after heat shock treatment in a thermotolerance test. In addition, transcription levels of all tested heat shock–related genes were significantly lower in the mutant than in the wild type. We also demonstrated that the mean lethal time to death (LT50) of ΔMrSVP significantly increased relative to the wild type in insect bioassays (both topical inoculation and injection) involving Galleria mellonella. Moreover, similar rates of appressorium formation between ΔMrSVP and the wild type—and the significantly different expression of virulence-related genes such as acid trehalase and sucrose nonfermenting protein kinase in the haemocoel after injection—revealed that MrSVP is required for virulence in the insect haemocoel. Conclusions Overall, our data suggest that the Mrsvp gene contributes to thermotolerance and virulence of M. robertsii. Furthermore, this gene is deeply involved in the mycosis of insect cadavers and in immune escape rather than insect cuticle penetration during infection

Petrogenesis and tectonic setting of the Shanzhuang monzogranites in central Jiangxi Province, South China: Evidence from lithology, geochemistry and zircon U-Pb geochronology

This paper presents a systemic study on geochronology, geochemistry and Sr-Nd isotopes of the northern monzogranite from Shanzhuang granitic pluton located on the southern part of the Qinhang Metallogenic Belt in central Jiangxi Province which comprises monzogranite and granodiorite to constrain its emplacement age, petrogenesis and tectonic setting. All zircons from the monzogranite are confirmed to be of magmatic origin by CL imaging, Th/U ratio and diagrams of (Sm/La)(N)-La and Ce/Ce*-(Sm/La)(N) and are divided into three groups. The weighted mean age (420 similar to 419Ma) of the first group is interpreted as crystallization age of the monzogranite. The Shanzhuang monzogranite, which has SiO2 content of 71.48% similar to 66.20%, K2O/Na2O ratio of 1.24 similar to 0.99 and A/CNK ratio of >1.1, with the emergences of muscovite and garnet as well as ACF diagram and negative correlations of Rb with Th and Y, belongs to high K calc-alkaline, strongly peraluminous S-type granite. Meanwhile, this type rock defined by a depletion in large ion lithophile elements Ba and Sr and high field strength elements Nb and Ta is interpreted as a product of partial melting of the crust-derived materials. The ratios of CaO/Na2O (0.14 similar to 0.07), Rb/Sr (8.27 similar to 3.83) and Rb/Ba (1.48 similar to 0.82), the content of FeOT + MgO + TiO2 (2.4 similar to 1.8) and the t(2DM) ages (2031 similar to 1971Ma) together suggest that the protolith of the monzogranite is Paleoproterozoie clay-rich argillaceous rocks. Moreover, the monzogranite is likely formed at a temperature condition (722 similar to 720 degrees C) constrained by saturation- and Ti temperatures of zircon, and a pressure condition of 2.71 similar to 3.21kbar constrained by the Ti-in-biotite geothermometer. Combined with the Paleozoic tectonic development of South China, the diagram of Rb-(Y + Nb) shows that the geological setting of Shanzhuang granitic pluton is post-collision. In summary, the Shanzhuang pluton was formed by partial melting of Paleoproterozoic clay-rich argillaceous rocks at a low temperature and a low pressure condition under post-collisional setting

Validation of reference genes for gene expression studies in virus-infected Nicotiana benthamiana using quantitative real-time PCR.

Nicotiana benthamiana is the most widely-used experimental host in plant virology. The recent release of the draft genome sequence for N. benthamiana consolidates its role as a model for plant-pathogen interactions. Quantitative real-time PCR (qPCR) is commonly employed for quantitative gene expression analysis. For valid qPCR analysis, accurate normalisation of gene expression against an appropriate internal control is required. Yet there has been little systematic investigation of reference gene stability in N. benthamiana under conditions of viral infections. In this study, the expression profiles of 16 commonly used housekeeping genes (GAPDH, 18S, EF1α, SAMD, L23, UK, PP2A, APR, UBI3, SAND, ACT, TUB, GBP, F-BOX, PPR and TIP41) were determined in N. benthamiana and those with acceptable expression levels were further selected for transcript stability analysis by qPCR of complementary DNA prepared from N. benthamiana leaf tissue infected with one of five RNA plant viruses (Tobacco necrosis virus A, Beet black scorch virus, Beet necrotic yellow vein virus, Barley stripe mosaic virus and Potato virus X). Gene stability was analysed in parallel by three commonly-used dedicated algorithms: geNorm, NormFinder and BestKeeper. Statistical analysis revealed that the PP2A, F-BOX and L23 genes were the most stable overall, and that the combination of these three genes was sufficient for accurate normalisation. In addition, the suitability of PP2A, F-BOX and L23 as reference genes was illustrated by expression-level analysis of AGO2 and RdR6 in virus-infected N. benthamiana leaves. This is the first study to systematically examine and evaluate the stability of different reference genes in N. benthamiana. Our results not only provide researchers studying these viruses a shortlist of potential housekeeping genes to use as normalisers for qPCR experiments, but should also guide the selection of appropriate reference genes for gene expression studies of N. benthamiana under other biotic and abiotic stress conditions

Characterization of transferrin receptor-mediated endocytosis and cellular iron delivery of recombinant human serum transferrin from rice (<it>Oryza sativa</it> L.)

Abstract Background Transferrin (TF) plays a critical physiological role in cellular iron delivery via the transferrin receptor (TFR)-mediated endocytosis pathway in nearly all eukaryotic organisms. Human serum TF (hTF) is extensively used as an iron-delivery vehicle in various mammalian cell cultures for production of therapeutic proteins, and is also being explored for use as a drug carrier to treat a number of diseases by employing its unique TFR-mediated endocytosis pathway. With the increasing concerns over the risk of transmission of infectious pathogenic agents of human plasma-derived TF, recombinant hTF is preferred to use for these applications. Here, we carry out comparative studies of the TFR binding, TFR-mediated endocytosis and cellular iron delivery of recombinant hTF from rice (rhTF), and evaluate its suitability for biopharmaceutical applications. Result Through a TFR competition binding affinity assay with HeLa human cervic carcinoma cells (CCL-2) and Caco-2 human colon carcinoma cells (HTB-37), we show that rhTF competes similarly as hTF to bind TFR, and both the TFR binding capacity and dissociation constant of rhTF are comparable to that of hTF. The endocytosis assay confirms that rhTF behaves similarly as hTF in the slow accumulation in enterocyte-like Caco-2 cells and the rapid recycling pathway in HeLa cells. The pulse-chase assay of rhTF in Caco-2 and HeLa cells further illustrates that rice-derived rhTF possesses the similar endocytosis and intracellular processing compared to hTF. The cell culture assays show that rhTF is functionally similar to hTF in the delivery of iron to two diverse mammalian cell lines, HL-60 human promyelocytic leukemia cells (CCL-240) and murine hybridoma cells derived from a Sp2/0-Ag14 myeloma fusion partner (HB-72), for supporting their proliferation, differentiation, and physiological function of antibody production. Conclusion The functional similarity between rice derived rhTF and native hTF in their cellular iron delivery, TFR binding, and TFR-mediated endocytosis and intracellular processing support that rice-derived rhTF can be used as a safe and animal-free alternative to serum hTF for bioprocessing and biopharmaceutical applications.</p

Identification of Glutathione Peroxidase (GPX) Gene Family in <i>Rhodiola crenulata</i> and Gene Expression Analysis under Stress Conditions

Glutathione peroxidases (GPXs) are important enzymes in the glutathione-ascorbate cycle for catalyzing the reduction of H2O2 or organic hydroperoxides to water. GPXs play an essential role in plant growth and development by participating in photosynthesis, respiration, and stress tolerance. Rhodiola crenulata is a popular traditional Chinese medicinal plant which displays an extreme energy of tolerance to harsh alpine climate. The GPXs gene family might provide R. crenulata for extensively tolerance to environment stimulus. In this study, five GPX genes were isolated from R. crenulata. The protein amino acid sequences were analyzed by bioinformation softwares with the results that RcGPXs gene sequences contained three conserve cysteine residues, and the subcellular location predication were in the chloroplast, endoplasmic reticulum, or cytoplasm. Five RcGPXs members presented spatial and temporal specific expression with higher levels in young and green organs. And the expression patterns of RcGPXs in response to stresses or plant hormones were investigated by quantitative real-time PCR. In addition, the putative interaction proteins of RcGPXs were obtained by yeast two-hybrid with the results that RcGPXs could physically interact with specific proteins of multiple pathways like transcription factor, calmodulin, thioredoxin, and abscisic acid signal pathway. These results showed the regulation mechanism of RcGPXs were complicated and they were necessary for R. crenulata to adapt to the treacherous weather in highland