Differential expression in leaves of Saccharum genotypes contrasting in biomass production provides evidence of genes involved in carbon partitioning

Background The development of biomass crops aims to meet industrial yield demands, in order to optimize profitability and sustainability. Achieving these goals in an energy crop like sugarcane relies on breeding for sucrose accumulation, fiber content and stalk number. To expand the understanding of the biological pathways related to these traits, we evaluated gene expression of two groups of genotypes contrasting in biomass composition. Results First visible dewlap leaves were collected from 12 genotypes, six per group, to perform RNA-Seq. We found a high number of differentially expressed genes, showing how hybridization in a complex polyploid system caused extensive modifications in genome functioning. We found evidence that differences in transposition and defense related genes may arise due to the complex nature of the polyploid Saccharum genomes. Genotypes within both biomass groups showed substantial variability in genes involved in photosynthesis. However, most genes coding for photosystem components or those coding for phosphoenolpyruvate carboxylases (PEPCs) were upregulated in the high biomass group. Sucrose synthase (SuSy) coding genes were upregulated in the low biomass group, showing that this enzyme class can be involved with sucrose synthesis in leaves, similarly to sucrose phosphate synthase (SPS) and sucrose phosphate phosphatase (SPP). Genes in pathways related to biosynthesis of cell wall components and expansins coding genes showed low average expression levels and were mostly upregulated in the high biomass group. Conclusions Together, these results show differences in carbohydrate synthesis and carbon partitioning in the source tissue of distinct phenotypic groups. Our data from sugarcane leaves revealed how hybridization in a complex polyploid system resulted in noticeably different transcriptomic profiles between contrasting genotypes.

Differential Expression in Leaves of Saccharum Genotypes Contrasting in Biomass Production Provides Evidence of Genes Involved in Carbon Partitioning

BackgroundThe development of biomass crops aims to meet industrial yield demands, in order to optimize profitability and sustainability. Achieving these goals in an energy crop like sugarcane relies on breeding for sucrose accumulation, fiber content and stalk number. To expand the understanding of the biological pathways related to these traits, we evaluated gene expression of two groups of genotypes contrasting in biomass composition.ResultsFirst visible dewlap leaves were collected from 12 genotypes, six per group, to perform RNA-Seq. We found a high number of differentially expressed genes, showing how hybridization in a complex polyploid system caused extensive modifications in genome functioning. We found evidence that differences in transposition and defense related genes may arise due to the complex nature of the polyploid Saccharum genomes. Genotypes within both biomass groups showed substantial variability in genes involved in photosynthesis. However, most genes coding for photosystem components or those coding for phosphoenolpyruvate carboxylases (PEPCs) were upregulated in the high biomass group. Sucrose synthase (SuSy) coding genes were upregulated in the low biomass group, showing that this enzyme class can be involved with sucrose synthesis in leaves, similarly to sucrose phosphate synthase (SPS) and sucrose phosphate phosphatase (SPP). Genes in pathways related to biosynthesis of cell wall components and expansins coding genes showed low average expression levels and were mostly upregulated in the high biomass group.ConclusionsTogether, these results show differences in carbohydrate synthesis and carbon partitioning in the source tissue of distinct phenotypic groups. Our data from sugarcane leaves revealed how hybridization in a complex polyploid system resulted in noticeably different transcriptomic profiles between contrasting genotypes.

Differential Expression in Leaves of Saccharum Genotypes Contrasting in Biomass Production Provides Evidence of Genes Involved in Carbon Partitioning

Background The development of biomass crops aims to meet industrial yield demands, in order to optimize profitability and sustainability. Achieving these goals in an energy crop like sugarcane relies on breeding for sucrose accumulation, fiber content and stalk number. To expand the understanding of the biological pathways related to these traits, we evaluated gene expression of two groups of genotypes contrasting in biomass composition. Results First visible dewlap leaves were collected from 12 genotypes, six per group, to perform RNA-SEq. We found a high number of differentially expressed genes, showing how hybridization in a complex polyploid system caused extensive modifications in genome functioning. We found evidence that the variation between these groups may be partly due to the expansion of the Saccharum genomes by differential transposition and defense related genes. Genotypes within both biomass groups showed substantial variability in genes involved in photosynthesis. However, most genes coding for photosystem components or those coding for phosphoenolpyruvate carboxylases (PEPCs) were upregulated in the high biomass group. Sucrose synthase (SuSy) coding genes were upregulated in the low biomass group, showing that this enzyme class can be involved with sucrose synthesis in leaves, similarly to sucrose phosphate synthase (SPS) and sucrose phosphate phosphatase (SPP). Genes in pathways related to biosynthesis of cell wall components and expansins coding genes showed low average expression levels and were mostly upregulated in the high biomass group. Conclusions Together, these results show differences in carbohydrate synthesis and carbon partitioning in the source tissue of distinct phenotypic groups. Our data from sugarcane leaves revealed how hybridization in a complex polyploid system resulted in noticeably different transcriptomic profiles between contrasting genotypes.

Tissue-autonomous phenylpropanoid production is essential for establishment of root barriers

ABSTRACTPlants deposit polymeric barriers in their root cell walls to protect against external stress and facilitate selective nutrient uptake. The compounds that make up these barriers originate from the fatty acid- and phenylpropanoid biosynthetic pathways. Although the machinery responsible for production of the barrier constituents is well-char-acterized, our pathway models lack spatiotemporal resolution – especially in roots - and the source tissue is often not clear due to the apoplastic nature of barriers. Insights into how the individual root tissues or cells contribute to forming apoplastic barriers is important for elucidation of their ultrastructure, function and development. Manipulation of the associated biosynthesis is delicate, as mutants often display pleiotropic phenotypes due to the broad role of the underlying metabolites. Here, we address these issues by creating a genetic tool that allows in vivo repression of the phenylpropanoid pathway with both spatial and temporal control. We provide strong evidence that tissue-auton-omous production of phenylpropanoids is essential for establishment of the endodermal Casparian strip. Moreover, we find that in order to maintain deposition and attachment of a coherent suberin matrix to the cell wall, cells require continuous production of aromatic constituents. This process is especially crucial in the suberized endodermis where we find that repression of phenylpropanoid production leads to active removal of suberin.

VULVAR TUMOR - CASE REPORT AND LITERATURE REVIEW

The vulvar region is a complex area because it comprises many elements, besides the skin itself. Therefore, it can present a variety of relatively rare tumors that can be classified based on source tissue (epithelial or mesenchymal). Benign epithelial tumors in the connective tissue are not often diagnosed in the vulvar area, which is mostly affected by fibroepithelial polyps.

Is repeated cypermethrin fumigation dangerous for the mitochondrial DNA in dry insect samples?

Entomological collections are the target of various insect pests, e.g. carpet beetles (Dermestidae) and booklice (Psocoptera) which can damage and completely destroy dry specimens in a relatively short time. Collections in the National Museum, Czech Republic (NMP) including the entomological ones are protected by fumigation using commercially available smoke shells ‘Cytrol Super SG’; fumigation is performed twice a year. The active insecticidal substance of these smoke shells is cypermethrin (6.25%). We tested whether the repeated cypermethrin fumigation of the NMP entomological collections negatively affects the quality of mitochondrial DNA in dry specimens and prevents the subsequent use of these samples for molecular analyses required for identification, taxonomy, systematics, and phylogenetic studies. We used 32 freshly fixed specimens of the flower chafer Oxythyrea funesta (Poda von Neuhaus, 1761) and 32 freshly fixed specimens of the brown-tailed cockroach Supella longipalpa (Fabricius, 1798). One half of specimens of both species was stored outside NMP and not fumigated (negative control), and the other half was deposited in collection hall with the NMP insect collection and directly exposed to the fumigation. Subsequently, all specimens were processed in a molecular laboratory under a standardized protocol using one leg as the source tissue after each fumigation, and the 658 bp long barcoding region of the cytochrome oxidase I (cox1) as the testing gene fragment. Results of the PCR product electrophoresis and the sequences acquired confirmed that the repeated fumigation had no negative effect on tested samples.

Cell-free DNA: the role in pathophysiology and as a biomarker in kidney diseases

Cell-free DNA (cfDNA) is present in various body fluids and originates mostly from blood cells. In specific conditions, circulating cfDNA might be derived from tumours, donor organs after transplantation or from the foetus during pregnancy. The analysis of cfDNA is mainly used for genetic analyses of the source tissue —tumour, foetus or for the early detection of graft rejection. It might serve also as a nonspecific biomarker of tissue damage in critical care medicine. In kidney diseases, cfDNA increases during haemodialysis and indicates cell damage. In patients with renal cell carcinoma, cfDNA in plasma and its integrity is studied for monitoring of tumour growth, the effects of chemotherapy and for prognosis. Urinary cfDNA is highly fragmented, but the technical hurdles can now be overcome and urinary cfDNA is being evaluated as a potential biomarker of renal injury and urinary tract tumours. Beyond its diagnostic application, cfDNA might also be involved in the pathogenesis of diseases affecting the kidneys as shown for systemic lupus, sepsis and some pregnancy-related pathologies. Recent data suggest that increased cfDNA is associated with acute kidney injury. In this review, we discuss the biological characteristics, sources of cfDNA, its potential use as a biomarker as well as its role in the pathogenesis of renal and urinary diseases.

Modulation of Immunoregulatory Properties of Mesenchymal Stromal Cells by Toll-Like Receptors: Potential Applications on GVHD

In the last decade, the immunomodulatory properties of mesenchymal stromal cells (MSCs) have attracted a lot of attention, due to their potential applicability in the treatment of graft-versus-host disease (GVHD), a condition frequently associated with opportunistic infections. The present review addresses how Pathogen-Associated Molecular Patterns (PAMPS) modulate the immunosuppressive phenotype of human MSCs by signaling through Toll-like receptors (TLRs). Overall, we observed that regardless of the source tissue, human MSCs express TLR2, TLR3, TLR4, and TLR9. Stimulation of distinct TLRs on MSCs elicits distinct inflammatory signaling pathways, differentially influencing the expression of inflammatory factors and the ability of MSCs to suppress the proliferation of immune system cells. The capacity to enhance the immunosuppressive phenotype of MSCs through TLRs stimulation might be properly elucidated in order to improve the MSC-based immunotherapy against GVHD.

Abstract 192: Axonal Guidance Factors as Potential Pediatric AVM Urinary Biomarker

Introduction: Development of noninvasive biomarkers for pediatric arteriovenous malformations (AVMs) could improve diagnosis and identify potential therapeutic targets. The axonal guidance factor, EphrinB2, and its cognate receptor EphB4 have been implicated in AVM development. Here we report our initial experience with urinary EphrinB2 as a novel AVM-specific urinary biomarker. Hypothesis: We hypothesize that urinary and tissue levels of EphrinB2 will be elevated in patients with AVM and distinguish AVM from other cerebrovascular disease. Methods: Specimens were collected from 45 pediatric patients (0-21 years of age) including unruptured AVM (n=15), moyamoya (n=15) and age- and sex-matched controls (n=15), per an IRB-approved protocol. ELISA quantified levels of urinary EphrinB2. Matched AVM tissue from the same patient was subjected to immunohistochemistry to correlate source tissue expression with biomarker levels. Results: Statistical analysis revealed a statistically significant increase in EphrinB2 and AVM-specific biomarker fingerprints capable of discerning AVM both from controls and also from other types of vascular neurosurgical disease. Compared to controls, EphrinB2 was elevated with mean EphrinB2 (0.10 pg/ug vs. 0.019 pg/ug, p<0.01) and compared to moyamoya patients mean EphrinB2 (0.10 pg/ug vs. 0.014, p<0.01). Similarly, EphrinB2 and EphB4 staining was significantly increased in AVM tissue vs. normal blood vessels matched as controls. Conclusion: In conclusion, these data suggests potential utility of urinary biomarkers, specifically EphrinB2, as diagnostic tools for clinicians treating children with AVMs and demonstrates proof-of- principle results supporting the concept of hypothesis-driven biomarker “fingerprinting” for unique cerebrovascular disease, differentiating AVM from other vascular pathology. For the first time, these data correlate known aspects of AVM molecular biology with clinically relevant diagnostics.