890 research outputs found
Role of Sex Peptide in Drosophila Males
Drosophila male sex peptide ACP70A is a small peptide mainly produced in the accessory glands. It elicits a high number of post-mating responses in mated females; yet its function in male physiology is not well known. Here, we explore its role in male sex behavior and pheromone biosynthesis, using males either mutant or RNAi knocked-down for Acp70A. Courtship was severely affected in both Acp70A mutants and Acp70A knocked-down males, with only 2% of the males succeeding copulation. Cuticular hydrocarbon amounts were moderately affected with 25% decrease in sp0 mutant (without Acp70A expression) and 10–22% increase in flies overexpressing Acp70A. Acp70A knock-down either ubiquitously or in the testes surprisingly resulted in an overproduction of hydrocarbons, whose amounts were double of the controls. We tested eight putative “off-target” genes but none of these led to an increase in hydrocarbon amounts. These results show that male courtship behavior is largely dependent on the presence of Acp70A and independent of cuticular hydrocarbons. The presence of potential “off-target” genes explaining the hydrocarbon phenotype is discussed
A Privacy-Aware Architecture for Demand Response Systems
We explore the privacy issues implicated by the development of demand response systems. We begin by highlighting the invasive nature of fine-granularity power consumption data, showing that the data collected by Advanced Metering Infrastructure (AMI) reveals detailed information about behavior within the home. We then show how privacy-aware design principles lead to novel system architectures that realize the benefits of demand response without requiring that AMI data be centrally collected. The resulting systems avoid both harm to subscribers and the potential need to scrap AMI-based demand response efforts in the face of public outcry. We also show that Trusted Platform Modules can be used to develop privacy-sensitive metering infrastructure
A survey of lineage-specific genes in Triticeae reveals de novo gene evolution from genomic raw material
Diploid plant genomes typically contain ~35,000 genes, almost all belonging to highly conserved gene families. Only a small fraction are lineage-specific, which are found in only one or few closely related species. Little is known about how genes arise de novo in plant genomes and how often this occurs; however, they are believed to be important for plants diversification and adaptation. We developed a pipeline to identify lineage-specific genes in Triticeae, using newly available genome assemblies of wheat, barley, and rye. Applying a set of stringent criteria, we identified 5942 candidate Triticeae-specific genes (TSGs), of which 2337 were validated as protein-coding genes in wheat. Differential gene expression analyses revealed that stress-induced wheat TSGs are strongly enriched in putative secreted proteins. Some were previously described to be involved in Triticeae non-host resistance and cold response. Additionally, we show that 1079 TSGs have sequence homology to transposable elements (TEs), ~68% of them deriving from regulatory non-coding regions of Gypsy retrotransposons. Most importantly, we demonstrate that these TSGs are enriched in transmembrane domains and are among the most highly expressed wheat genes overall. To summarize, we conclude that de novo gene formation is relatively rare and that Triticeae probably possess ~779 lineage-specific genes per haploid genome. TSGs, which respond to pathogen and environmental stresses, may be interesting candidates for future targeted resistance breeding in Triticeae. Finally, we propose that non-coding regions of TEs might provide important genetic raw material for the functional innovation of TM domains and the evolution of novel secreted proteins
Grasshopper Lazarillo, a GPI-anchored Lipocalin, increases Drosophila longevity and stress resistance, and functionally replaces its secreted homolog NLaz
Producción CientíficaLazarillo (Laz) is a glycosyl-phosphatidylinositol (GPI)-linked glycoprotein first characterized in the
developing nervous system of the grasshopper Schistocerca americana. It belongs to the Lipocalins,
a functionally diverse family of mostly secreted proteins. In this work we test whether the protective
capacity known for Laz homologs in flies and vertebrates (NLaz, GLaz and ApoD) is evolutionarily
conserved in grasshopper Laz, and can be exerted from the plasma membrane in a cell-autonomous
manner. First we demonstrate that extracellular forms of Laz have autocrine and paracrine protecting
effects for oxidative stress-challenged Drosophila S2 cells. Then we assay the effects of overexpressing
GPI-linked Laz in adult Drosophila and whether it rescues both known and novel phenotypes of NLaz null
mutants. Local effects of GPI-linked Laz inside and outside the nervous system promote survival upon
different stress forms, and extend lifespan and healthspan of the flies in a cell-type dependent manner.
Outside the nervous system, expression in fat body cells but not in hemocytes results in protection.
Within the nervous system, glial cell expression is more effective than neuronal expression. Laz actions
are sexually dimorphic in some expression domains. Fat storage promotion and not modifications in
hydrocarbon profiles or quantities explain the starvationedesiccation resistance caused by Laz overexpression.
This effect is exerted when Laz is expressed ubiquitously or in dopaminergic cells, but not in
hemocytes. Grasshopper Laz functionally restores the loss of NLaz, rescuing stress-sensitivity as well as
premature accumulation of aging-related damage, monitored by advanced glycation end products
(AGEs). However Laz does not rescue NLaz courtship behavioral defects. Finally, the presence of two new
Lipocalins with predicted GPI-anchors in mosquitoes shows that the functional advantages of
GPI-linkage have been commonly exploited by Lipocalins in the arthropodan lineage
Wheat gene bank accessions as a source of new alleles of the powdery mildew resistance gene Pm3: a large scale allele mining project
BACKGROUND: In the last hundred years, the development of improved wheat cultivars has led to the replacement of landraces and traditional varieties by modern cultivars. This has resulted in a decline in the genetic diversity of agriculturally used wheat. However, the diversity lost in the elite material is somewhat preserved in crop gene banks. Therefore, the gene bank accessions provide the basis for genetic improvement of crops for specific traits and and represent rich sources of novel allelic variation.
RESULTS: We have undertaken large scale molecular allele mining to isolate new alleles of the powdery mildew resistance gene Pm3 from wheat gene bank accessions. The search for new Pm3 alleles was carried out on a geographically diverse set of 733 wheat accessions originating from 20 countries. Pm3 specific molecular tools as well as classical pathogenicity tests were used to characterize the accessions. Two new functional Pm3 alleles were identified out of the eight newly cloned Pm3 sequences. These new resistance alleles were isolated from accessions from China and Nepal. Thus, the repertoire of functional Pm3 alleles now includes 17 genes, making it one of the largest allelic series of plant resistance genes. The combined information on resistant and susceptible Pm3 sequences will allow to study molecular function and specificity of functional Pm3 alleles.
CONCLUSIONS: This study demonstrates that molecular allele mining on geographically defined accessions is a useful strategy to rapidly characterize the diversity of gene bank accessions at a specific genetic locus of agronomical importance. The identified wheat accessions with new resistance specificities can be used for marker-assisted transfer of the Pm3 alleles to modern wheat lines
454 sequencing put to the test using the complex genome of barley
BACKGROUND: During the past decade, Sanger sequencing has been used to completely sequence hundreds of microbial and a few higher eukaryote genomes. In recent years, a number of alternative technologies became available, among them adaptations of the pyrosequencing procedure (i.e. "454 sequencing"), promising a ~100-fold increase in throughput over Sanger technology – an advancement which is needed to make large and complex genomes more amenable to full genome sequencing at affordable costs. Although several studies have demonstrated its potential usefulness for sequencing small and compact microbial genomes, it was unclear how the new technology would perform in large and highly repetitive genomes such as those of wheat or barley. RESULTS: To study its performance in complex genomes, we used 454 technology to sequence four barley Bacterial Artificial Chromosome (BAC) clones and compared the results to those from ABI-Sanger sequencing. All gene containing regions were covered efficiently and at high quality with 454 sequencing whereas repetitive sequences were more problematic with 454 sequencing than with ABI-Sanger sequencing. 454 sequencing provided a much more even coverage of the BAC clones than ABI-Sanger sequencing, resulting in almost complete assembly of all genic sequences even at only 9 to 10-fold coverage. To obtain highly advanced working draft sequences for the BACs, we developed a strategy to assemble large parts of the BAC sequences by combining comparative genomics, detailed repeat analysis and use of low-quality reads from 454 sequencing. Additionally, we describe an approach of including small numbers of ABI-Sanger sequences to produce hybrid assemblies to partly compensate the short read length of 454 sequences. CONCLUSION: Our data indicate that 454 pyrosequencing allows rapid and cost-effective sequencing of the gene-containing portions of large and complex genomes and that its combination with ABI-Sanger sequencing and targeted sequence analysis can result in large regions of high-quality finished genomic sequences
Distribution, functional impact, and origin mechanisms of copy number variation in the barley genome
BACKGROUND There is growing evidence for the prevalence of copy number variation (CNV) and its role in phenotypic variation in many eukaryotic species. Here we use array comparative genomic hybridization to explore the extent of this type of structural variation in domesticated barley cultivars and wild barleys. RESULTS A collection of 14 barley genotypes including eight cultivars and six wild barleys were used for comparative genomic hybridization. CNV affects 14.9% of all the sequences that were assessed. Higher levels of CNV diversity are present in the wild accessions relative to cultivated barley. CNVs are enriched near the ends of all chromosomes except 4H, which exhibits the lowest frequency of CNVs. CNV affects 9.5% of the coding sequences represented on the array and the genes affected by CNV are enriched for sequences annotated as disease-resistance proteins and protein kinases. Sequence-based comparisons of CNV between cultivars Barke and Morex provided evidence that DNA repair mechanisms of double-strand breaks via single-stranded annealing and synthesis-dependent strand annealing play an important role in the origin of CNV in barley. CONCLUSIONS We present the first catalog of CNVs in a diploid Triticeae species, which opens the door for future genome diversity research in a tribe that comprises the economically important cereal species wheat, barley, and rye. Our findings constitute a valuable resource for the identification of CNV affecting genes of agronomic importance. We also identify potential mechanisms that can generate variation in copy number in plant genomes.This work was financially supported by the following grants:
project GABI-BARLEX, German Federal Ministry of Education and Research
(BMBF), #0314000 to MP, US, KFXM and NS; Triticeae Coordinated
Agricultural Project, USDA-NIFA #2011-68002-30029 to GJM; and Agriculture
and Food Research Initiative Plant Genome, Genetics and Breeding Program
of USDA’s Cooperative State Research and Extension Service, #2009-65300-
05645 to GJM
BITC Sensitizes Pancreatic Adenocarcinomas to TRAIL-Induced Apoptosis
Pancreatic adenocarcinoma is an aggressive cancer with a greater than 95% mortality rate and short survival after diagnosis. Chemotherapeutic resistance hinders successful treatment. This resistance is often associated with mutations in codon 12 of the K-Ras gene (K-Ras 12), which is present in over 90% of all pancreatic adenocarcinomas. Codon 12 mutations maintain Ras in a constitutively active state leading to continuous cellular proliferation. Our study determined if TRAIL resistance in pancreatic adenocarcinomas with K-Ras 12 mutations could be overcome by first sensitizing the cells with Benzyl isothiocyanate (BITC). BITC is a component of cruciferous vegetables and a cell cycle inhibitor. BxPC3, MiaPaCa2 and Panc-1 human pancreatic adenocarcinoma cell lines were examined for TRAIL resistance. Our studies show BITC induced TRAIL sensitization by dual activation of both the extrinsic and intrinsic apoptotic pathways
Wheat zinc finger protein TaZF interacts with both the powdery mildew AvrPm2 protein and the corresponding wheat Pm2a immune receptor
Plant defense responses to pathogens are induced after direct or indirect perception of effector proteins or their activity on host proteins. In fungal-plant interactions, relatively little is known about whether, in addition to avirulence effectors and immune receptors, other proteins contribute to specific recognition. The nucleotide-binding leucine-rich repeat (NLR) immune receptor Pm2a in wheat recognizes the fungal powdery mildew effector AvrPm2. We found that the predicted wheat zinc finger TaZF interacts with both the fungal avirulence protein AvrPm2 and the wheat NLR Pm2a. We further demonstrated that the virulent AvrPm2-H2 variant does not interact with TaZF. TaZF silencing in wheat resulted in a reduction but not a loss of Pm2a-mediated powdery mildew resistance. Interaction studies showed that the leucine-rich repeat domain of Pm2a is the mediator of the interaction with TaZF. TaZF recruits both Pm2a and AvrPm2 from the cytosol to the nucleus, resulting in nuclear localization of Pm2a, TaZF, and AvrPm2 in wheat. We propose that TaZF acts as a facilitator of Pm2a-dependent AvrPm2 effector recognition. Our findings highlight the importance of identifying effector host targets for characterization of NLR-mediated effector recognition
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