44 Resistenzzüchtung / Widerstandsfähigkeit gegen Schadorganismen I

44-1 - Bewertung der Feldresistenz verschiedener Winterrapssorten gegenüber Verticillium longisporum mittels quantitativer PCRClassification of winter oilseed rape resistance towards the soilborne pathogen Verticillium longisporum by quantitative PCRJessica Knüfer, Daniel Teshome Lopisso, Birger Koopmann, Andreas von Tiedemann44-2 - Identification and characterization of three putative compatibility factor genes involved in the plant – Verticillium interactionIdentifikation und Charakterisierung drei putativer Kompatibilitätsfaktoren in der Pflanze – Verticillium InteraktionRoxana Hossain, Lisa Krapoth, Dirk Schenke, Daguang Cai44-3 - Impact of cultivar resistance to Verticillium longisporum on drought stress tolerance of winter oilseed rape (Brassica napus)Einfluss der Sortenresistenz gegen Verticillium longisporum auf die Trockenstresstoleranz von Winterraps (Brassica napus)Daniel Lopisso, Jessica Knüfer, Birger Koopmann, Andreas von Tiedemann44-4 - Wirksamkeit von Majorgenen in Raps gegenüber Phoma lingam unter Berücksichtigung steigender Temperaturen und des PathotypenspektrumsEfficacy of major genes in oilseed rape against Phoma lingam with regard to rising temperatures and the population structureMark Winter, Coretta Klöppel , Fadeke Fajemisin, Birger Koopmann44-5 - Anfälligkeit von Raps -Resynthesen und -Sorten auf den Rapsstängelrüssler (Ceutorhynchus napi Gyll.) Befall – potentielle ResistenzfaktorenSusceptibility of resynthesized lines and cultivars of oilseed rape on rape stem weevil (Ceutorhynchus napi Gyll.) infestation – potential plant traits responsible for resistanceHeike Schäfer-Kösterke, Bernd Ulber44-6 - Zweijähriges Rassen-Monitoring von Exserohilum turcicum in europäischen MaisanbaugebietenTwo-year race monitoring for Exserohilum turcicum in European maize growing regionsHendrik Hanekamp, Andreas von Tiedemann, Birger Koopmann44-7 - Smart breeding und Nutzung des Genpools von Wildarten zur Verbesserung der Krankheitsresistenz von KartoffelnSmart breeding and exploitation of the genepool from wild species for the improvement of disease resistance in potatoJanine König, Marion Nachtigall, Ramona Thieme, Jörg Schubert44-8 - Neue Ansätze für eine effizientere Resistenzzüchtung bei RebenNew approaches for increasing efficieny of grapevine resistance breedingRudolf Eibach, Reinhard Töpfe

Comparative genomics of Australian isolates of the wheat stem rust pathogen Puccinia graminis f. sp. tritici reveals extensive polymorphism in candidate effector genes

The wheat stem rust fungus Puccinia graminis f. sp. tritici (Pgt) is one of the most destructive pathogens of wheat. In this study, a draft genome was built for a founder Australian Pgt isolate of pathotype (pt.) 21-0 (collected in 1954) by next generation DNA sequencing. A combination of reference-based assembly using the genome of the previously sequenced American Pgt isolate CDL 75-36-700-3 (p7a) and de novo assembly were performed resulting in a 92 Mbp reference genome for Pgt isolate 21-0. Approximately 13 Mbp of de novo assembled sequence in this genome is not present in the p7a reference assembly. This novel sequence is not specific to 21-0 as it is also present in three other Pgt rust isolates of independent origin. The new reference genome was subsequently used to build a pan-genome based on five Australian Pgt isolates. Transcriptomes from germinated urediniospores and haustoria were separately assembled for pt. 21-0 and comparison of gene expression profiles showed differential expression in ∼10% of the genes each in germinated spores and haustoria. A total of 1,924 secreted proteins were predicted from the 21-0 transcriptome, of which 520 were classified as haustorial secreted proteins (HSPs). Comparison of 21-0 with two presumed clonal field derivatives of this lineage (collected in 1982 and 1984) that had evolved virulence on four additional resistance genes (Sr5, Sr11, Sr27, SrSatu) identified mutations in 25 HSP effector candidates. Some of these mutations could explain their novel virulence phenotypes.Authors wish to thank the Two Blades Foundation for financial support. Part of this work was supported through access to facilities managed by Bioplatforms Australia and funded by the Australian Government National Collaborative Research Infrastructure Strategy and Education Investment Fund Super Science Initiative

A Novel Device for the Measurement of the Mechanical and Magnetic Axes of Superconducting Magnet Assemblies for Accelerators

In the context of the LHC superconducting magnet production, especially for dipoles and quadrupoles due to their complexity, it is foreseen to perform acceptance tests, at an early production stage, to detect possible significant deviations from the design values. The knowledge of the magnetic field geometry is very important, especially for the main magnets. In order to get this information a new device has been conceived that measures the magnets at room temperature during different stages of construction. This device incorporates a sensitive measuring probe and an efficient data acquisition system because the coils are only powered at about 10-5 of the nominal D.C. current. It is dedicated to Quadrupole and Dipole (by using Quadrupole-Configured Dipole (QCD) transformation) magnets, but is also easily adaptable to higher order magnets (n = 3, 4 and 5) by specific orientation of the search coils. It is equipped with magnetic sensors (4 fixed tangential coils and AC excitation current for the magnet) and position sensors (3D-laser tracker and light reflector) that allow the simultaneous detection of the magnetic field axis and the cold bore axis. It is equipped as well with a set of 4 LEDs and associated with a CCD camera that allows both the measurement of the cold bore diameter and its position with respect to the mole. This paper describes the system and reports the first results measured on the pre-series magnets recently assembled

The cell adhesion molecule L1 regulates the expression of choline acetyltransferase and the development of septal cholinergic neurons

Mutations in the L1 gene cause severe brain malformations and mental retardation. We investigated the potential roles of L1 in the regulation of choline acetyltransferase (ChAT) and in the development of septal cholinergic neurons, which are known to project to the hippocampus and play key roles in cognitive functions. Using stereological approaches, we detected significantly fewer ChAT-positive cholinergic neurons in the medial septum and vertical limb of the diagonal band of Broca (MS/VDB) of 2-week-old L1-deficient mice compared to wild-type littermates (1644 ± 137 vs. 2051 ± 165, P = 0.038). ChAT protein levels in the septum were 53% lower in 2-week-old L1-deficient mice compared to wild-type littermates. ChAT activity in the septum was significantly reduced in L1-deficient mice compared to wild-type littermates at 1 (34%) and 2 (40%) weeks of age. In vitro, increasing doses of L1-Fc induced ChAT activity in septal neurons with a significant linear trend (*P = 0.0065). At 4 weeks of age in the septum and at all time points investigated in the caudate-putamen (CPu), the number of ChAT-positive neurons and the levels of ChAT activity were not statistically different between L1-deficient mice and wild-type littermates. The total number of cells positive for the neuronal nuclear antigen (NeuN) in the MS/VDB and CPu was not statistically different in L1-deficient mice compared to wild-type littermates, and comparable expression of the cell cycle marker Ki67 was observed. Our results indicate that L1 is required for the timely maturation of septal cholinergic neurons and that L1 promotes the expression and activity of ChAT in septal neurons

The tomato Prf complex is a molecular trap for bacterial effectors based on Pto transphosphorylation

The bacteria Pseudomonas syringae is a pathogen of many crop species and one of the model pathogens for studying plant and bacterial arms race coevolution. In the current model, plants perceive bacteria pathogens via plasma membrane receptors, and recognition leads to the activation of general defenses. In turn, bacteria inject proteins called effectors into the plant cell to prevent the activation of immune responses. AvrPto and AvrPtoB are two such proteins that inhibit multiple plant kinases. The tomato plant has reacted to these effectors by the evolution of a cytoplasmic resistance complex. This complex is compromised of two proteins, Prf and Pto kinase, and is capable of recognizing the effector proteins. How the Pto kinase is able to avoid inhibition by the effector proteins is currently unknown. Our data shows how the tomato plant utilizes dimerization of resistance proteins to gain advantage over the faster evolving bacterial pathogen. Here we illustrate that oligomerisation of Prf brings into proximity two Pto kinases allowing them to avoid inhibition by the effectors by transphosphorylation and to activate immune responses

Mol Vis

PURPOSE: To analyze in vivo the function of chicken acidic leucine-rich epidermal growth factor-like domain containing brain protein/Neuroglycan C (gene symbol: Cspg5) during retinal degeneration in the Rpe65(-)/(-) mouse model of Leber congenital amaurosis. METHODS: We resorted to mice with targeted deletions in the Cspg5 and retinal pigment epithelium protein of 65 kDa (Rpe65) genes (Cspg5(-)/(-)/Rpe65(-)/(-)). Cone degeneration was assessed with cone-specific peanut agglutinin staining. Transcriptional expression of rhodopsin (Rho), S-opsin (Opn1sw), M-opsin (Opn1mw), rod transducin alpha subunit (Gnat1), and cone transducin alpha subunit (Gnat2) genes was assessed with quantitative PCR from 2 weeks to 12 months. The retinal pigment epithelium (RPE) was analyzed at P14 with immunodetection of the retinol-binding protein membrane receptor Stra6. RESULTS: No differences in the progression of retinal degeneration were observed between the Rpe65(-)/(-) and Cspg5(-)/(-)/Rpe65(-)/(-) mice. No retinal phenotype was detected in the late postnatal and adult Cspg5(-)/(-) mice, when compared to the wild-type mice. CONCLUSIONS: Despite the previously reported upregulation of Cspg5 during retinal degeneration in Rpe65(-)/(-) mice, no protective effect or any involvement of Cspg5 in disease progression was identified

Master singular behavior for the Sugden factor of the one-component fluids near their gas-liquid critical point

We present the master (i.e. unique) behavior of the squared capillary length - so called the Sudgen factor-, as a function of the temperature-like field along the critical isochore, asymptotically close to the gas-liquid critical point of twenty (one component) fluids. This master behavior is obtained using the scale dilatation of the relevant physical fields of the one-component fluids. The scale dilatation introduces the fluid-dependent scale factors in a manner analog with the linear relations between physical fields and scaling fields needed by the renormalization theory applied to the Ising-like universality class. The master behavior for the Sudgen factor satisfies hyperscaling and can be asymptotically fitted by the leading terms of the theoretical crossover functions for the correlation length and the susceptibility in the homogeneous domain recently obtained from massive renormalization in field theory. In the absence of corresponding estimation of the theoretical crossover functions for the interfacial tension, we define the range of the temperature-like field where the master leading power law can be practically used to predict the singular behavior of the Sudgen factor in conformity with the theoretical description provided by the massive renormalization scheme within the extended asymptotic domain of the one-component fluid "subclass"

The Genomic Analysis of Erythrocyte microRNA Expression in Sickle Cell Diseases

BACKGROUND: Since mature erythrocytes are terminally differentiated cells without nuclei and organelles, it is commonly thought that they do not contain nucleic acids. In this study, we have re-examined this issue by analyzing the transcriptome of a purified population of human mature erythrocytes from individuals with normal hemoglobin (HbAA) and homozygous sickle cell disease (HbSS). METHODS AND FINDINGS: Using a combination of microarray analysis, real-time RT-PCR and Northern blots, we found that mature erythrocytes, while lacking ribosomal and large-sized RNAs, contain abundant and diverse microRNAs. MicroRNA expression of erythrocytes was different from that of reticulocytes and leukocytes, and contributed the majority of the microRNA expression in whole blood. When we used microRNA microarrays to analyze erythrocytes from HbAA and HbSS individuals, we noted a dramatic difference in their microRNA expression pattern. We found that miR-320 played an important role for the down-regulation of its target gene, CD71 during reticulocyte terminal differentiation. Further investigation revealed that poor expression of miR-320 in HbSS cells was associated with their defective downregulation CD71 during terminal differentiation. CONCLUSIONS: In summary, we have discovered significant microRNA expression in human mature erythrocytes, which is dramatically altered in HbSS erythrocytes and their defect in terminal differentiation. Thus, the global analysis of microRNA expression in circulating erythrocytes can provide mechanistic insights into the disease phenotypes of erythrocyte diseases

A Genetic Screen Reveals Arabidopsis Stomatal and/or Apoplastic Defenses against Pseudomonas syringae pv. tomato DC3000

Bacterial infection of plants often begins with colonization of the plant surface, followed by entry into the plant through wounds and natural openings (such as stomata), multiplication in the intercellular space (apoplast) of the infected tissues, and dissemination of bacteria to other plants. Historically, most studies assess bacterial infection based on final outcomes of disease and/or pathogen growth using whole infected tissues; few studies have genetically distinguished the contribution of different host cell types in response to an infection. The phytotoxin coronatine (COR) is produced by several pathovars of Pseudomonas syringae. COR-deficient mutants of P. s. tomato (Pst) DC3000 are severely compromised in virulence, especially when inoculated onto the plant surface. We report here a genetic screen to identify Arabidopsis mutants that could rescue the virulence of COR-deficient mutant bacteria. Among the susceptible to coronatine-deficient Pst DC3000 (scord) mutants were two that were defective in stomatal closure response, two that were defective in apoplast defense, and four that were defective in both stomatal and apoplast defense. Isolation of these three classes of mutants suggests that stomatal and apoplastic defenses are integrated in plants, but are genetically separable, and that COR is important for Pst DC3000 to overcome both stomatal guard cell- and apoplastic mesophyll cell-based defenses. Of the six mutants defective in bacterium-triggered stomatal closure, three are defective in salicylic acid (SA)-induced stomatal closure, but exhibit normal stomatal closure in response to abscisic acid (ABA), and scord7 is compromised in both SA- and ABA-induced stomatal closure. We have cloned SCORD3, which is required for salicylic acid (SA) biosynthesis, and SCORD5, which encodes an ATP-binding cassette (ABC) protein, AtGCN20/AtABCF3, predicted to be involved in stress-associated protein translation control. Identification of SCORD5 begins to implicate an important role of stress-associated protein translation in stomatal guard cell signaling in response to microbe-associated molecular patterns and bacterial infection

Embryonic Stem Cell-Derived L1 Overexpressing Neural Aggregates Enhance Recovery after Spinal Cord Injury in Mice

An obstacle to early stem cell transplantation into the acutely injured spinal cord is poor survival of transplanted cells. Transplantation of embryonic stem cells as substrate adherent embryonic stem cell-derived neural aggregates (SENAs) consisting mainly of neurons and radial glial cells has been shown to enhance survival of grafted cells in the injured mouse brain. In the attempt to promote the beneficial function of these SENAs, murine embryonic stem cells constitutively overexpressing the neural cell adhesion molecule L1 which favors axonal growth and survival of grafted and imperiled cells in the inhibitory environment of the adult mammalian central nervous system were differentiated into SENAs and transplanted into the spinal cord three days after compression lesion. Mice transplanted with L1 overexpressing SENAs showed improved locomotor function when compared to mice injected with wild-type SENAs. L1 overexpressing SENAs showed an increased number of surviving cells, enhanced neuronal differentiation and reduced glial differentiation after transplantation when compared to SENAs not engineered to overexpress L1. Furthermore, L1 overexpressing SENAs rescued imperiled host motoneurons and parvalbumin-positive interneurons and increased numbers of catecholaminergic nerve fibers distal to the lesion. In addition to encouraging the use of embryonic stem cells for early therapy after spinal cord injury L1 overexpression in the microenvironment of the lesioned spinal cord is a novel finding in its functions that would make it more attractive for pre-clinical studies in spinal cord regeneration and most likely other diseases of the nervous system