Studies on Peroxisome Motility in the Model Fungal System Ustilago maydis

Peroxisomes are ubiquitous organelles found in almost all eukaryotes. They are sensitive to changes in cellular homeostasis and involved in various metabolic processes. Deficiencies in peroxisome function cause severe neurological problems. Here I report, investigation of peroxisome motility and its relation to peroxisomal functions in the fungal model system Ustilago maydis. Peroxisomes are mostly motile in Ustilago maydis. Motile peroxisomes show different motility patterns: short-range pulse type movements and long range bidirectional motility. Motility behaviour is not static as oscillating peroxisomes may start long-range motility. Here, I present evidence that long-range bidirectional peroxisome motility is an energy driven process and is essential for homogeneous distribution of peroxisomes. Similar to early endosomes and endoplasmic reticulum, microtubule motors kinesin-3 and dynein are responsible for long-range peroxisome transport. In addition to using the same molecular motors for transport, early endosomes, endoplasmic reticulum and peroxisomes have the same transport velocity. Interestingly, motile peroxisomes and endoplasmic reticulum tubules co-localize with early endosomes. Functional investigation of early endosome mutants, Δrab5a and Yup1ts has revealed a novel transport mechanism where endoplasmic reticulum and peroxisomes hitch hike on early endosomes. Additionally, I report functional characterization of an AAA-ATPase, um05592, which has high homology to human protein NP_055873. Altogether these results reveal molecular mechanism of peroxisome transport in Ustilago maydis. Similarities in transport machinery illustrate Ustilago maydis as a model system to study peroxisome function in mammalian cells

Comparative analysis of plant immune receptor architectures uncovers host proteins likely targeted by pathogens.

BACKGROUND: Plants deploy immune receptors to detect pathogen-derived molecules and initiate defense responses. Intracellular plant immune receptors called nucleotide-binding leucine-rich repeat (NLR) proteins contain a central nucleotide-binding (NB) domain followed by a series of leucine-rich repeats (LRRs), and are key initiators of plant defense responses. However, recent studies demonstrated that NLRs with non-canonical domain architectures play an important role in plant immunity. These composite immune receptors are thought to arise from fusions between NLRs and additional domains that serve as "baits" for the pathogen-derived effector proteins, thus enabling pathogen recognition. Several names have been proposed to describe these proteins, including "integrated decoys" and "integrated sensors". We adopt and argue for "integrated domains" or NLR-IDs, which describes the product of the fusion without assigning a universal mode of action. RESULTS: We have scanned available plant genome sequences for the full spectrum of NLR-IDs to evaluate the diversity of integrations of potential sensor/decoy domains across flowering plants, including 19 crop species. We manually curated wheat and brassicas and experimentally validated a subset of NLR-IDs in wild and cultivated wheat varieties. We have examined NLR fusions that occur in multiple plant families and identified that some domains show re-occurring integration across lineages. Domains fused to NLRs overlap with previously identified pathogen targets confirming that they act as baits for the pathogen. While some of the integrated domains have been previously implicated in disease resistance, others provide new targets for engineering durable resistance to plant pathogens. CONCLUSIONS: We have built a robust reproducible pipeline for detecting variable domain architectures in plant immune receptors across species. We hypothesize that NLR-IDs that we revealed provide clues to the host proteins targeted by pathogens, and that this information can be deployed to discover new sources of disease resistance

In planta determination of GaMyb transcription factor as a target of pathogen induced microRNA, mir159

One of the molecular mechanisms underlying the plant–pathogen interactions is the regulation of gene expressions in plant responses by microRNAs which are either stimulated or silenced against pathogen attacks. Among the plant miRNAs, we found that mir159 is one of which that differentially expressed upon Blumeria graminis f. sp. hordei (Bgh) infected resistant and susceptible barley lines. The study aims to confirm its role in regulating one of its putative target genes, GaMyb transcription factor, in vivo. Thus, both mir159 and GaMyb genes were co-infiltrated into tobacco as constructs of pEarlyGate100 and GFP expressing pEarlyGate103, respectively, via agrobacterium transient transformation. Our results suggest that mir159 indeed reduces the expression of GaMyb transcription factor by which, for the first time, confirm it being a biological target of mir159. To further confirm the induced expression level and the biological role of mir159 in susceptible barley by virulent Bgh pathogen infection, the differential level of disease development should be investigated in a susceptible line of barley after the over expression and/or the silencing of the gene of mir159

A new ZTL-type F-box functions as a positive regulator in disease resistance: VIGS analysis in barley against powdery mildew

In our previous studies, we found a new Zeitlupe (ZTL) type F-box protein which is expressed at a higher level upon avirulent pathogen infection (Bozkurt et al., 2007). F-box proteins mark the proteins to be degraded through 26S proteasome system by ubiquitination. Since the information on the role of ubiquitin mediated proteolysis in disease responses is advancing rapidly, we sought to understand the way which F-box functions in resistance response as part of ubiquitin-proteasome pathway. Interestingly, in response to silencing of this F-box gene via BSMV mediated virus induced gene silencing (VIGS) method, barley plants lost resistance towards avirulent pathogen race. The Pallas-01 line having Mla1 R-gene showed hyphae formations when inoculated with avirulent powdery mildew race, Bgh103, after 4-fold silencing. This observation suggests that F-box protein functions as a positive regulator in powdery mildew disease mechanism and broadens the function of ZTL-type F-box proteins, previously known to have roles only in circadian clocks, flowering time control, and phytochrome pathway

Septin-mediated plant cell invasion by the rice blast fungus, Magnaporthe oryzae

Author's accepted version, please cite the published version by following the DOI link.To cause rice blast disease, the fungus Magnaporthe oryzae develops a pressurized dome-shaped cell called an appressorium, which physically ruptures the leaf cuticle to gain entry to plant tissue. Here, we report that a toroidal F-actin network assembles in the appressorium by means of four septin guanosine triphosphatases, which polymerize into a dynamic, hetero-oligomeric ring. Septins scaffold F-actin, via the ezrin-radixin-moesin protein Tea1, and phosphatidylinositide interactions at the appressorium plasma membrane. The septin ring assembles in a Cdc42- and Chm1-dependent manner and forms a diffusion barrier to localize the inverse-bin-amphiphysin-RVS-domain protein Rvs167 and the Wiskott-Aldrich syndrome protein Las17 at the point of penetration. Septins thereby provide the cortical rigidity and membrane curvature necessary for protrusion of a rigid penetration peg to breach the leaf surface.Biotechnology and Biological Sciences Research Council (BBSRC)ER

Septin-mediated plant cell invasion by the rice blast fungus, Magnaporthe oryzae

Blasting Through The fungus that causes rice blast disease, Magnaporthe oryzae , can lead to devastating reductions in rice yields. M. oryzae enters the plant by developing specialized infection structures called appressoria. Appressoria generate enormous internal turgor pressure that somehow creates invasive forces that physically break the plant cuticle. Dagdas et al. (p. 1590 ) found that a toroidal (doughnut-shaped) filamentous actin network forms at the base of the appressorium at the precise point where the penetration peg, which ruptures the rice leaf cuticle, will emerge. This network is scaffolded by means of four septin guanosine triphosphatases, which form a dynamic ring structure that colocalizes with F-actin. The findings reveal how fungi translate extreme pressure into localized physical force. </jats:p

Use of mobilization and felaxation techniques in patients with periarthritis humeroscapulatis

T. aestivum fusion validation by RNA-seq. Summary of manual validation of T. aestivum NLR-IDs using RNA-seq data. (XLSX 60 kb

Additional file 8: of Comparative analysis of plant immune receptor architectures uncovers host proteins likely targeted by pathogens

Manual verification of domains predicted by high-throughput scripts with webservers for brassica, tomato, wheat and soybean. (XLSX 22 kb