Characterisation of the legume SERK-NIK gene superfamily including splice variants: Implications for development and defence

<p>Abstract</p> <p>Background</p> <p><it>SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE </it>(<it>SERK</it>) genes are part of the regulation of diverse signalling events in plants. Current evidence shows SERK proteins function both in developmental and defence signalling pathways, which occur in response to both peptide and steroid ligands. <it>SERKs </it>are generally present as small gene families in plants, with five <it>SERK </it>genes in Arabidopsis. Knowledge gained primarily through work on Arabidopsis SERKs indicates that these proteins probably interact with a wide range of other receptor kinases and form a fundamental part of many essential signalling pathways. The <it>SERK1 </it>gene of the model legume, <it>Medicago truncatula </it>functions in somatic and zygotic embryogenesis, and during many phases of plant development, including nodule and lateral root formation. However, other <it>SERK </it>genes in <it>M. truncatula </it>and other legumes are largely unidentified and their functions unknown.</p> <p>Results</p> <p>To aid the understanding of signalling pathways in <it>M. truncatula</it>, we have identified and annotated the <it>SERK </it>genes in this species. Using degenerate PCR and database mining, eight more <it>SERK</it>-like genes have been identified and these have been shown to be expressed. The amplification and sequencing of several different PCR products from one of these genes is consistent with the presence of splice variants. Four of the eight additional genes identified are upregulated in cultured leaf tissue grown on embryogenic medium. The sequence information obtained from <it>M. truncatula </it>was used to identify <it>SERK </it>family genes in the recently sequenced soybean (<it>Glycine max</it>) genome.</p> <p>Conclusions</p> <p>A total of nine <it>SERK </it>or <it>SERK-like </it>genes have been identified in <it>M. truncatula </it>and potentially 17 in soybean. Five <it>M. truncatula SERK </it>genes arose from duplication events not evident in soybean and Lotus. The presence of splice variants has not been previously reported in a <it>SERK </it>gene. Upregulation of four newly identified <it>SERK </it>genes (in addition to the previously described <it>MtSERK1</it>) in embryogenic tissue cultures suggests these genes also play a role in the process of somatic embryogenesis. The phylogenetic relationship of members of the <it>SERK </it>gene family to closely related genes, and to development and defence function is discussed.</p

The 2HA line of Medicago truncatula has characteristics of an epigenetic mutant that is weakly ethylene insensitive

BACKGROUND The Medicago truncatula 2HA seed line is highly embryogenic while the parental line Jemalong rarely produces embryos. The 2HA line was developed from one of the rare Jemalong regenerates and this method for obtaining a highly regenerable genotype in M. truncatula is readily reproducible suggesting an epigenetic mechanism. Microarray transcriptomic analysis showed down regulation of an ETHYLENE INSENSITIVE 3-like gene in 2HA callus which provided an approach to investigating epigenetic regulation of genes related to ethylene signalling and the 2HA phenotype. Ethylene is involved in many developmental processes including somatic embryogenesis (SE) and is associated with stress responses. RESULTS Microarray transcriptomic analysis showed a significant number of up-regulated transcripts in 2HA tissue culture, including nodule and embryo specific genes and transposon-like genes, while only a few genes were down-regulated, including an EIN3-like gene we called MtEIL1. This reduced expression was associated with ethylene insensitivity of 2HA plants that was further investigated. The weak ethylene insensitivity affected root and nodule development. Sequencing of MtEIL1 found no difference between 2HA and wild-type plants. DNA methylation analysis of MtEIL1 revealed significant difference between 2HA and wild-type plants. Tiling arrays demonstrated an elevated level of miRNA in 2HA plants that hybridised to the antisense strand of the MtEIL1 gene. AFLP-like methylation profiling revealed more differences in DNA methylation between 2HA and wild-type. Segregation analysis demonstrated the recessive nature of the eil1 phenotype and the dominant nature of the SE trait. CONCLUSIONS We have demonstrated that EIL1 of Medicago truncatula (MtEIL1) is epigenetically silenced in the 2HA seed line. The possible cause is an elevated level of miRNA that targets its 3'UTR and is also associated with DNA methylation of MtEIL1. Down regulation of MtEIL1 makes it possible to form nodules in the presence of ethylene and affects root growth under normal conditions. Segregation analysis showed no association between MtEIL1 expression and SE in culture but the role and mechanism of ethylene signalling in the process of plant regeneration through SE requires further investigation. The work also suggests that epigenetic changes to a particular gene induced in culture can be fixed in regenerated plants.This work was funded by the Australian Research Council (CEO348212) through the ARC Centre of Excellence for Integrative Legume Research (CILR)

High Passage MIN6 Cells Have Impaired Insulin Secretion with Impaired Glucose and Lipid Oxidation

Type 2 diabetes is a metabolic disorder characterized by the inability of beta-cells to secrete enough insulin to maintain glucose homeostasis. MIN6 cells secrete insulin in response to glucose and other secretagogues, but high passage (HP) MIN6 cells lose their ability to secrete insulin in response to glucose. We hypothesized that metabolism of glucose and lipids were defective in HP MIN6 cells causing impaired glucose stimulated insulin secretion (GSIS). HP MIN6 cells had no first phase and impaired second phase GSIS indicative of global functional impairment. This was coupled with a markedly reduced ATP content at basal and glucose stimulated states. Glucose uptake and oxidation were higher at basal glucose but ATP content failed to increase with glucose. HP MIN6 cells had decreased basal lipid oxidation. This was accompanied by reduced expressions of Glut1, Gck, Pfk, Srebp1c, Ucp2, Sirt3, Nampt. MIN6 cells represent an important model of beta cells which, as passage numbers increased lost first phase but retained partial second phase GSIS, similar to patients early in type 2 diabetes onset. We believe a number of gene expression changes occurred to produce this defect, with emphasis on Sirt3 and Nampt, two genes that have been implicated in maintenance of glucose homeostasis.These authors have no support or funding to report

Effects of N-functional groups on the electron transfer kinetics of VO²⁺/VO₂⁺ at carbon:Decoupling morphology from chemical effects using model systems

Carbons and nanocarbons are important electrode materials for vanadium redox flow battery applications, however, the kinetics of vanadium species are often sluggish at these surfaces, thus prompting interest in functionalization strategies to improve performance. Herein, we investigate the effect of N-functionalities on the VO2+/VO2+ redox process at carbon electrodes. We fabricate thin film carbon disk electrodes that are metal-free, possess well-defined geometry and display smooth topography, while featuring different N-site distribution, thus enabling a mechanistic investigation of the intrinsic surface activity towards VO2+/VO2+. Voltammetry and electrochemical impedance spectroscopy show that N-functionalities improve performance, with pyridinic/pyrrolic-N imparting the most significant improvements in charge transfer rates and reversibility, compared to graphitic-N. This was further supported by voltammetry studies on nitrogen-free electrodes modified via aryldiazonium chemistry with molecular pyridyl adlayers. Computational modeling using an electrochemical-chemical mechanism indicates that introduction of surface pyridinic/pyrrolic-N can increase the heterogeneous rate constants by approximately two orders of magnitude relative to those observed at nitrogen-free carbon (k0 = 1.29 × 10−4 vs 9.34 × 10−7 cm/s). Simulations also suggest that these N-functionalities play a role in affecting reaction rates in the chemical step. Our results indicate that nitrogen incorporation via basic functional groups offers an interesting route to the design of advanced carbon electrodes for VRFB devices.</p

Speech rhythm: a metaphor?

Is speech rhythmic? In the absence of evidence for a traditional view that languages strive to coordinate either syllables or stress-feet with regular time intervals, we consider the alternative that languages exhibit contrastive rhythm subsisting merely in the alternation of stronger and weaker elements. This is initially plausible, particularly for languages with a steep ‘prominence gradient’, i.e. a large disparity between stronger and weaker elements; but we point out that alternation is poorly achieved even by a ‘stress-timed’ language such as English, and, historically, languages have conspicuously failed to adopt simple phonological remedies that would ensure alternation. Languages seem more concerned to allow ‘syntagmatic contrast’ between successive units and to use durational effects to support linguistic functions than to facilitate rhythm. Furthermore, some languages (e.g. Tamil, Korean) lack the lexical prominence which would most straightforwardly underpin prominence alternation. We conclude that speech is not incontestibly rhythmic, and may even be antirhythmic. However, its linguistic structure and patterning allow the metaphorical extension of rhythm in varying degrees and in different ways depending on the language, and that it is this analogical process which allows speech to be matched to external rhythms

Characterizing Cosmic-Ray Propagation in Massive Star-forming Regions: The Case of 30 Doradus and the Large Magellanic Cloud

Using infrared, radio, and γ-ray data, we investigate the propagation characteristics of cosmic-ray (CR) electrons and nuclei in the 30 Doradus (30 Dor) star-forming region in the Large Magellanic Cloud (LMC) using a phenomenological model based on the radio-far-infrared correlation within galaxies. Employing a correlation analysis, we derive an average propagation length of ~100-140 pc for ~3 GeV CR electrons resident in 30 Dor from consideration of the radio and infrared data. Assuming that the observed γ-ray emission toward 30 Dor is associated with the star-forming region, and applying the same methodology to the infrared and γ-ray data, we estimate a ~20 GeV propagation length of 200-320 pc for the CR nuclei. This is approximately twice as large as for ~3 GeV CR electrons, corresponding to a spatial diffusion coefficient that is ~4 times higher, scaling as (R/GV)δ with δ ≈ 0.7-0.8 depending on the smearing kernel used in the correlation analysis. This value is in agreement with the results found by extending the correlation analysis to include ~70 GeV CR nuclei traced by the 3-10 GeV γ-ray data (δ ≈ 0.66 ± 0.23). Using the mean age of the stellar populations in 30 Dor and the results from our correlation analysis, we estimate a diffusion coefficient D_R ≈ (0.9-1.0) × 10^(27)(R/GV)0.7 cm^(2) s^(–1). We compare the values of the CR electron propagation length and surface brightness for 30 Dor and the LMC as a whole with those of entire disk galaxies. We find that the trend of decreasing average CR propagation distance with increasing disk-averaged star formation activity holds for the LMC, and extends down to single star-forming regions, at least for the case of 30 Dor

The stellar content of low redshift BL Lac host galaxies from multicolour imaging

We present B-band imaging of 18 low redshift (z<0.3) BL Lac objects for which their host galaxies were previously resolved in the R-band and the near-infrared H-band. For a subset of the objects, also U- and V-band imaging is presented. These data are used to investigate the blue-red-near-infrared colours and the colour gradients of the host galaxies of BL Lacs in comparison with other elliptical galaxies with and without nuclear activity. In all cases galaxies are well represented by an elliptical model, with average absolute magnitude M_B=-21.6+-0.7 and average scale length R_e=7.6+-3.2 kpc. The best-fit B-band Kormendy relation is in reasonable agreement with that obtained for normal ellipticals and radio galaxies. This structural and dynamical similarity indicates that all massive elliptical galaxies can experience nuclear activity without significant perturbation of their global structure. The distributions of the integrated blue/near-infrared colour (with average B-H=3.5+-0.5) and colour gradient (with average Delta(B-R)/Delta(log r)=-0.14+-0.75) of the BL Lac hosts are much wider than those for normal ellipticals, and most BL Lac objects have bluer hosts and/or steeper colour gradients than those in normal ellipticals. The blue colours are likely caused by a young stellar population component, and indicates a link between star formation caused by an interaction/merging event and the onset of the nuclear activity. This result is corroborated by stellar population modelling, indicating a presence of young/intermediate age populations in the majority of the sample, in agreement with low redshift quasar hosts. The lack of strong signs of interaction may require a significant time delay between the event with associated star formation episodes and the start of the nuclear activity.Comment: 16 pages, 10 figure

First Steps towards Underdominant Genetic Transformation of Insect Populations

The idea of introducing genetic modifications into wild populations of insects to stop them from spreading diseases is more than 40 years old. Synthetic disease refractory genes have been successfully generated for mosquito vectors of dengue fever and human malaria. Equally important is the development of population transformation systems to drive and maintain disease refractory genes at high frequency in populations. We demonstrate an underdominant population transformation system in Drosophila melanogaster that has the property of being both spatially self-limiting and reversible to the original genetic state. Both population transformation and its reversal can be largely achieved within as few as 5 generations. The described genetic construct {Ud} is composed of two genes; (1) a UAS-RpL14.dsRNA targeting RNAi to a haploinsufficient gene RpL14 and (2) an RNAi insensitive RpL14 rescue. In this proof-of-principle system the UAS-RpL14.dsRNA knock-down gene is placed under the control of an Actin5c-GAL4 driver located on a different chromosome to the {Ud} insert. This configuration would not be effective in wild populations without incorporating the Actin5c-GAL4 driver as part of the {Ud} construct (or replacing the UAS promoter with an appropriate direct promoter). It is however anticipated that the approach that underlies this underdominant system could potentially be applied to a number of species. Figure

X-ray observations of three young, early-type galaxies

Massive haloes of hot plasma exist around some, but not all elliptical galaxies. There is evidence that this is related to the age of the galaxy. In this paper, new X-ray observations are presented for three early-type galaxies that show evidence of youth, in order to investigate their X-ray components and properties. NGC 5363 and NGC 2865 were found to have X-ray emission dominated by purely discrete stellar sources. Limits are set on the mass distribution in one of the galaxies observed with XMM–Newton, NGC 4382, which contains significant hot gas. We detect the X-ray emission in NGC 4382 out to 4re. The mass-to-light ratio is consistent with a stellar origin in the inner regions but rises steadily to values indicative of some dark matter by 4re. These results are set in context with other data drawn from the literature, for galaxies with ages estimated from dynamical or spectroscopic indicators. Ages obtained from optical spectroscopy represent central luminosity-weighted stellar ages. We examine the X-ray evolution with age, normalized by B- and K-band luminosities. Low values of Log(LX/LB) and Log(LX/LK) are found for all galaxies with ages between 1 and 4 Gyr. Luminous X-ray emission only appears in older galaxies. This suggests that the interstellar medium is removed and then it takes several gigayears for hot gas haloes to build up, following a merger. A possible mechanism for gas expulsion might be associated with feedback from an active nucleus triggered during a merger