Unraveling the senses of Phytophthora; leads to novel control strategies?

Oomycetes cause devastating diseases on plants and animals. They cause major yield losses in many crop plants and their control heavily depends on agrochemicals. This is certainly true for the potato late blight pathogen Phytophthora infestans. Strong concerns about adverse effects of agrochemicals on food safety and environment are incentives for the development of novel, environmental friendly control strategies preferably based on natural products. Cyclic lipopeptides (CLPs) were recently discovered as a new class of natural compounds with strong activities against oomycetes including Phytophthora. CLPs lyse zoospores, inhibit mycelial growth and effectively reduce late blight disease. In order to unravel how Phytophthora senses CLPs and other environmental signals we follow two approaches. On the one hand, we monitor genome wide changes in gene expression induced by CLPs with the aim to identify the cellular pathways targeted by CLPs. On the other hand, we analyse components of ubiquitous signal transduction pathways with the aim to identify features that are unique for Phytophthora or oomycetes and, hence, could be suitable targets for novel anti-oomycete agents. Mining and comparing whole genome sequences have revealed that Phytophthora harbours many novel phospholipid modifying enzymes, unique for oomycetes. They have aberrant combinations of catalytic and regulatory domains occasionally combined with transmembrane domains. The latter resemble receptors that might be activated by extracellular ligands. Phospholipids, the substrates of these enzymes, are structural membrane components that also function in signalling. Together these findings open new avenues of research aimed at target-discovery in oomycetes

The aspartic proteinase family of three Phytophthora species

Background - Phytophthora species are oomycete plant pathogens with such major social and economic impact that genome sequences have been determined for Phytophthora infestans, P. sojae and P. ramorum. Pepsin-like aspartic proteinases (APs) are produced in a wide variety of species (from bacteria to humans) and contain conserved motifs and landmark residues. APs fulfil critical roles in infectious organisms and their host cells. Annotation of Phytophthora APs would provide invaluable information for studies into their roles in the physiology of Phytophthora species and interactions with their hosts. Results - Genomes of Phytophthora infestans, P. sojae and P. ramorum contain 11-12 genes encoding APs. Nine of the original gene models in the P. infestans database and several in P. sojae and P. ramorum (three and four, respectively) were erroneous. Gene models were corrected on the basis of EST data, consistent positioning of introns between orthologues and conservation of hallmark motifs. Phylogenetic analysis resolved the Phytophthora APs into 5 clades. Of the 12 sub-families, several contained an unconventional architecture, as they either lacked a signal peptide or a propart region. Remarkably, almost all APs are predicted to be membrane-bound. Conclusions - One of the twelve Phytophthora APs is an unprecedented fusion protein with a putative G-protein coupled receptor as the C-terminal partner. The others appear to be related to well-documented enzymes from other species, including a vacuolar enzyme that is encoded in every fungal genome sequenced to date. Unexpectedly, however, the oomycetes were found to have both active and probably-inactive forms of an AP similar to vertebrate BACE, the enzyme responsible for initiating the processing cascade that generates the Aß peptide central to Alzheimer's Disease. The oomycetes also encode enzymes similar to plasmepsin V, a membrane-bound AP that cleaves effector proteins of the malaria parasite Plasmodium falciparum during their translocation into the host red blood cell. Since the translocation of Phytophthora effector proteins is currently a topic of intense research activity, the identification in Phytophthora of potential functional homologues of plasmepsin V would appear worthy of investigation. Indeed, elucidation of the physiological roles of the APs identified here offers areas for future study. The significant revision of gene models and detailed annotation presented here should significantly facilitate experimental design

Acclimation to salt modifies the activation of several osmotic stress-activated lipid signalling pathways in <em>Chlamydomonas</em>

Osmotic stress rapidly activates several phospholipid signalling pathways in the unicellular alga Chlamydomonas. In this report, we have studied the effects of salt-acclimation on growth and phospholipid signalling. Growing cells on media containing 100 mM NaCl increased their salt-tolerance but did not affect the overall phospholipid content, except that levels of phosphatidylinositol phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] were reduced by one-third. When these NaCl-acclimated cells were treated with increasing concentrations of salt, the same lipid signalling pathways as in non-acclimated cells were activated. This was witnessed as increases in phosphatidic acid (PA), lyso-phosphatidic acid (L-PA), diacylglycerol pyrophosphate (DGPP), PI(4,5)P2 and its isomer PI(3,5)P2. However, all dose-dependent responses were shifted to higher osmotic-stress levels, and the responses were lower than in non-acclimated cells. When NaCl-acclimated cells were treated with other osmotica, such as KCl and sucrose, the same effects were found, illustrating that they were due to hyperosmotic rather than hyperionic acclimation. The results indicate that acclimation to moderate salt stress modifies stress perception and the activation of several downstream pathways

Phytophthora-genomics: nieuwe mogelijkheden en uitdagingen

Het onderzoek aan de ziekteverwekker Phytophthora infestans is in een stroomversnelling geraakt. Dit is grotendeels te danken aan de stormachtige ontwikkelingen in genomics en de nieuwe inzichten die uit het genoomonderzoek voortvloeien. De DNA-code van het genoom van Phytophthora infestans en van enkele verwante soorten is ontrafeld en via internet beschikbaar. We staan pas aan het begin van het ontginnen van een schat aan gegevens die in de genoomsequentie verborgen ligt maar er zijn nu al verrassende vindingen gedaan die kunnen leiden tot nieuwe strategiëen voor de bestrijding van de aardappelziekte. De moleculaire basis van de 'gen-om-gen'-interactie tussen aardappel en Phytophthora infestans wordt steeds duidelijker en we kunnen, op basis van geconserveerde motieven in effectoreiwitten, genen aanwijzen die een rol spelen in gastheerspecifiteit. Nu kunnen we bestaande theoriën gaan staven met experimentele bewijzen en voorspellingen doen over de duurzaamheid van resistentiegenen. We hebben genen gevonden die coderen voor eiwitten die niet voorkomen in andere organismen en daarom mogelijk interessante aangrijpingspunten zijn voor nieuwe Phytophthora-bestrijdingsmiddelen of een ingang bieden voor alternatieve vormen van bestrijding. Er is nu onomstotelijk bewijs dat Phytophthora, ondanks zijn schimmelachtige uiterlijk, niet tot schimmelrijk behoort maar groene voorouders heeft die over fotosynthetiserende organellen beschikte

Samenvattingen Fusarium

An overview has been made on the theme of Fusarium with short summaries of research projects. Various topics are discussed. The titles of the summaries are: Dissection of lineage specific chromosomes in the onion pathogen Fusarium oxysporum f. sp. cepae and other emerging UK pathosystems; Differences in pre-harvest practices linked with the occurrence of Fusarium species and associated mycotoxins in maize from two ethnic groups in Vietnam; Biology, control and detection of Fusarium diseases in UK horticulture; Fusarium poae reduces Fusarium graminearum infection and mycotoxin production; European Union Reference Laboratory for mycotoxins & plant toxins in food and feed; Global epidemic of Panama disease on banana is caused by a new fungal species originating from Southeast Asia; Epidemiology of Fusarium in greenhouses

Consensus molecular subtype classification of colorectal adenomas

Consensus molecular subtyping is an RNA expression-based classification system for colorectal cancer (CRC). Genomic alterations accumulate during CRC pathogenesis, including the premalignant adenoma stage, leading to changes in RNA expression. Only a minority of adenomas progress to malignancies, a transition that is associated with specific DNA copy number aberrations or microsatellite instability (MSI). We aimed to investigate whether colorectal adenomas can already be stratified into consensus molecular subtype (CMS) classes, and whether specific CMS classes are related to the presence of specific DNA copy number aberrations associated with progression to malignancy. RNA sequencing was performed on 62 adenomas and 59 CRCs. MSI status was determined with polymerase chain reaction-based methodology. DNA copy number was assessed by low-coverage DNA sequencing (n = 30) or array-comparative genomic hybridisation (n = 32). Adenomas were classified into CMS classes together with CRCs from the study cohort and from The Cancer Genome Atlas (n = 556), by use of the established CMS classifier. As a result, 54 of 62 (87%) adenomas were classified according to the CMS. The CMS3 ‘metabolic subtype’, which was least common among CRCs, was most prevalent among adenomas (n = 45; 73%). One of the two adenomas showing MSI was classified as CMS1 (2%), the ‘MSI immune’ subtype. Eight adenomas (13%) were classified as the ‘canonical’ CMS2. No adenomas were classified as the ‘mesenchymal’ CMS4, consistent with the fact that adenomas lack invasion-associated stroma. The distribution of the CMS classes among adenomas was confirmed in an independent series. CMS3 was enriched with adenomas at low risk of progressing to CRC, whereas relatively more high-risk adenomas were observed in CMS2. We conclude that adenomas can be stratified into the CMS classes. Considering that CMS1 and CMS2 expression signatures may mark adenomas at increased risk of progression, the distribution of the CMS classes among adenomas is consistent with the proportion of adenomas expected to progress to CRC