Heterologous Expression of Secreted Proteins from Phytophthora in Pichia pastoris Followed by Protein Purification by Immobilized Metal Ion Affinity (IMAC).

This chapter describes the protocol for heterologous expression of Phytophthora proteins in the yeast Pichia pastoris. Two methods to prepare the constructs for expression are described, using two different strains of P. pastoris, as well as methods for protein expression and purification by immobilized metal ion affinity (IMAC).

A Method for the Separation of Phytophthora Oosporesfrom Soil for DNA-Based Detection.

Here, we present a protocol for the isolation and detection of Phytophthora oospores directly from soil samples. Our method incorporates a novel technique for isolating Phytophthora oospores using filter pouches and an improved DNA extraction procedure specifically designed for oospores. While we have primarily developed this protocol for detecting P.

A draft genome sequence of sp. ICMP 17802, isolated from root nodules of .

Here, we report the draft genome sequence of sp. ICMP 17802. This bacterium was obtained from the International Collection of Microorganisms from Plants and was originally isolated from root nodules of .

Draft genome sequence of a endosymbiont from (Fritsch, 1958) (Acari, Syringophilidae).

We present the draft genome of a endosymbiont from quill mites. This is the first representative of a recently discovered distinct lineage (supergroup P). We hope the genome will be a useful resource for comparative evolutionary and genomic studies across the globally distributed symbiont .

[Preoperative fasting in children-Experiences with implementing a liberal fasting policy].

Background: Traditional fasting rules for children prior to elective operations ("6‑4‑2 rule") often lead to prolonged fasting times with possible adverse events (discomfort, hypoglycemia, metabolic disorders, agitation/delirium). A new liberal fasting policy allowing children to drink clear fluids until being called to the operating room ("6‑4‑0") was established in our university hospital. This article describes our experiences and retrospectively examines the effects.

Plant-pathogen management in a native forest ecosystem.

Forest ecosystems all over the world are facing a growing threat from plant-disease outbreaks. As pollution, climate change, and global pathogen movement intensify, so too do the impacts of forest pathogens. In this essay, we examine a case study of the New Zealand kauri tree (Agathis australis) and its oomycetepathogen, Phytophthora agathidicida.

Harnessing CRISPR-Cas for oomycete genome editing.

Oomycetes are a group of microorganisms that include pathogens responsible for devastating diseases in plants and animals worldwide. Despite their importance, the development of genome editing techniques for oomycetes has progressed more slowly than for model microorganisms. Here, we review recent breakthroughs in clustered regularly interspaced short palindromic repeats (CRISPR)-Cas technologies that are expanding the genome editing toolbox for oomycetes - from the original Cas9 study to Cas12a editing, ribonucleoprotein (RNP) delivery, and complementation.

Rapid divergence in independent aspects of the compatibility phenotype in a interaction.

Heritable symbionts represent important components of the biology, ecology and evolution of their arthropod hosts. Particular microbial taxa have become common across arthropods as a consequence of their ability to establish in new host species. For a host shift to occur, the symbiont must be exposed to a novel host and then be compatible: it must not cause excess pathology, must have good vertical transmission and must possess a drive phenotype that enables spread.

Evaluation of a Clinical Decision Support System for the most evidence-based approach to managing perioperative anticoagulation.

Study Objective: We explored the feasibility of a Clinical Decision Support System (CDSS) to guide evidence-based perioperative anticoagulation.

Design: Prospective randomised clinical management simulation multicentre study.

Setting: Five University and 11 general hospitals in Germany.

Disruption of KCC2 in Parvalbumin-Positive Interneurons Is Associated With a Decreased Seizure Threshold and a Progressive Loss of Parvalbumin-Positive Interneurons.

GABA receptors are ligand-gated ion channels, which are predominantly permeable for chloride. The neuronal K-Cl cotransporter KCC2 lowers the intraneuronal chloride concentration and thus plays an important role for GABA signaling. KCC2 loss-of-function is associated with seizures and epilepsy.

Microbial protection favors parasite tolerance and alters host-parasite coevolutionary dynamics.

Coevolution between hosts and parasites is a major driver of rapid evolutionary change and diversification. However, direct antagonistic interactions between hosts and parasites could be disrupted when host microbiota form a line of defense, a phenomenon widespread across animal and plant species. By suppressing parasite infection, protective microbiota could reduce the need for host-based defenses and favor host support for microbiota colonization, raising the possibility that the microbiota can alter host-parasite coevolutionary patterns and processes.

Assessing the effectiveness of oxathiapiprolin toward , the causal agent of kauri dieback disease.

species cause disease and devastation of plants in ecological and horticultural settings worldwide. A recently identified species, , infects and ultimately kills the treasured kauri trees () that are endemic to New Zealand. Currently, there are few options for managing kauri dieback disease.

The Fatty Acid Methyl Ester (FAME) profile of Phytophthora agathidicida and its potential use as diagnostic tool.

Phytophthora diseases cause devastation to crops and native ecosystems worldwide. In New Zealand, Phytophthora agathidicida is threatening the survival of kauri, an endemic, culturally and ecologically important tree species. The current method for detecting P.

Evidence for multiple colonisations and Wolbachia infections shaping the genetic structure of the widespread butterfly Polyommatus icarus in the British Isles.

The paradigm of isolation in southern refugia during glacial periods followed by expansions during interglacials, producing limited genetic differentiation in northern areas, dominates European phylogeography. However, the existence of complex structured populations in formerly glaciated areas, and islands connected to mainland areas during glacial maxima, call for alternative explanations. We reconstructed the mtDNA phylogeography of the widespread Polyommatus Icarus butterfly with an emphasis on the formerly glaciated and connected British Isles.

Complete Genome Sequence of a Novel Pseudomonas fluorescens Strain Isolated from the Flower of Kūmarahou (Pomaderris kumeraho).

Kūmarahou (Pomaderris kumeraho) is a shrub endemic to New Zealand used in rongoā (traditional medicine). While studying the antimicrobial properties of kūmarahou, we isolated a new strain of Pseudomonas fluorescens, which we designated KF1 (for "kūmarahou flower 1"). Here, we report the complete genome sequence of P.

A single point mutation converts a glutaryl-7-aminocephalosporanic acid acylase into an N-acyl-homoserine lactone acylase.

Objective: To change the specificity of a glutaryl-7-aminocephalosporanic acid acylase (GCA) towards N-acyl homoserine lactones (AHLs; quorum sensing signalling molecules) by site-directed mutagenesis.

Results: Seven residues were identified by analysis of existing crystal structures as potential determinants of substrate specificity. Site-saturation mutagenesis libraries were created for each of the seven selected positions.

Screening a Natural Product-Inspired Library for Anti- Activities.

is a genus of microorganisms that cause devastating dieback and root-rot diseases in thousands of plant hosts worldwide. The economic impact of diseases on crops and native ecosystems is estimated to be billions of dollars per annum. These invasive pathogens are extremely difficult to control using existing chemical means, and the effectiveness of the few treatments available is being jeopardized by increasing rates of resistance.

Structure of a double CACHE chemoreceptor ligand-binding domain from Pseudomonas syringae provides insights into the basis of proline recognition.

Chemotaxis is the process of sensing chemical gradients and navigating towards favourable conditions. Bacterial chemotaxis is mediated by arrays of trans-membrane chemoreceptor proteins. The most common class of chemoreceptors have periplasmic ligand-binding domains (LBDs) that detect extracellular chemical signs and transduce these signals to the downstream chemotaxis machinery.

Host genotype and genetic diversity shape the evolution of a novel bacterial infection.

Pathogens continue to emerge from increased contact with novel host species. Whilst these hosts can represent distinct environments for pathogens, the impacts of host genetic background on how a pathogen evolves post-emergence are unclear. In a novel interaction, we experimentally evolved a pathogen (Staphylococcus aureus) in populations of wild nematodes (Caenorhabditis elegans) to test whether host genotype and genetic diversity affect pathogen evolution.

Rapid molecular evolution of symbionts of .

is a genus of whose members include plant pathogens, insect pathogens and endosymbionts of animals. phenotypes have been repeatedly observed to be spontaneously lost in cultures, and several studies have documented a high genomic turnover in symbionts and plant pathogens. These observations suggest that evolves quickly in comparison to other insect symbionts.

Transcript Variants Have Distinct Roles in Ovarian Carcinoma and Differently Influence Platinum Sensitivity and Angiogenesis.

The prognosis of late-stage epithelial ovarian cancer (EOC) patients is affected by chemotherapy response and the malignant potential of the tumor cells. In earlier work, we identified hypermethylation of the gene () as a prognostic biomarker and contrary functions of transcript variants (TV1 and TV2) in A2780 and SKOV3 cells. The aim of the study was to further validate these results and to increase the knowledge about function in EOC.

Infection Patterns and Fitness Effects of and Symbionts in the Green Lacewing .

Endosymbionts are widely distributed in insects and can strongly affect their host ecology. The common green lacewing () is a neuropteran insect which is widely used in biological pest control. However, their endosymbionts and their interactions with their hosts have not been very well studied.

Discordant Supramolecular Fibres Reversibly Depolymerised by Temperature and Light.

Synthetic stimuli responsive supramolecular polymers attract increasing interest for their ability to mimic the unique properties of natural assemblies. Here we focus on the well-studied benzene-1,3,5-tricarboxamide (BTA) motif, and substitute it with two (S)-3,7-dimethyloctyl groups and an azobenzene photoswitch. We demonstrate the UV (λ=365 nm) induced depolymerisation of the helical hydrogen-bonded polymers in methylcyclohexane (MCH) through circular dichroism and UV-vis spectroscopy in dilute solution (15 μm), and NMR and iPAINT super-resolution microscopy in concentrated solution (300 μm).