Dynamische ziekteverwekkers; wat we (willen) weten over oömyceten

Inaugurele rede bij de aanvaarding van het ambt van persoonlijk hoogleraar bij het Laboratorium voor Fytopathologie aan Wageningen Universiteit op 11 juni 2009: "Dynamische ziekteverwekkers : Wat we (willen) weten over oömyceten". In deze rede over oömyceten wordt ingegaan op de plaats van deze ziekteverwekker in de evolutie. Daarna op de oömoceet Phytophthora infestans, de verwekker van de aardappelziekte, ziektebestrijding en resistentieveredeling, de diversiteit van Phytophthora en zijn gastheren en de genetische blauwdruk: het DNA, de genen, de effectoren en de resistentiegene

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

Avirulentiegenen in Phytophthora infestans, de veroorzaker van de aardappelziekte

Een proefschrift van Wageningen Universiteit beschrijft het karteren en kloneren van Avr-genen in Phytophthora infestans. Ook is de fenotypische en genotypische diversiteit in P. infestans-veldisolaten in Noord China onderzoch

Internuclear gene silencing in Phytophthora infestans is established through chromatin remodelling

In the plant pathogen Phytophthora infestans, nuclear integration of inf1 transgenic DNA sequences results in internuclear gene silencing of inf1. Although silencing is regulated at the transcriptional level, it also affects transcription from other nuclei within heterokaryotic cells of the mycelium. Here we report experiments exploring the mechanism of internuclear gene silencing in P. infestans. The DNA methylation inhibitor 5-azacytidine induced reversion of the inf1-silenced state. Also, the histone deacetylase inhibitor trichostatin-A was able to reverse inf1 silencing. inf1-expression levels returned to the silenced state when the inhibitors were removed except in non-transgenic inf1-silenced strains that were generated via internuclear gene silencing, where inf1 expression was restored permanently. Therefore, inf1-transgenic sequences are required to maintain the silenced state. Prolonged culture of non-transgenic inf1-silenced strains resulted in gradual reactivation of inf1 gene expression. Nuclease digestion of inf1-silenced and non-silenced nuclei showed that inf1 sequences in silenced nuclei were less rapidly degraded than non-silenced inf1 sequences. Bisulfite sequencing of the endogenous inf1 locus did not result in detection of any cytosine methylation. Our findings suggest that the inf1-silenced state is based on chromatin remodelling

Biologically active Phytophthora mating hormone prepared by catalytic asymmetric total synthesis

A Phytophthora mating hormone with an array of 1,5-stereogenic centers has been synthesized by using our recently developed methodology of catalytic enantioselective conjugate addition of Grignard reagents. We applied this methodology in a diastereo- and enantioselective iterative route and obtained two of the 16 possible stereoisomers of Phytophthora hormone α1. These synthetic stereoisomers induced the formation of sexual spores (oospores) in A2 mating type strains of three heterothallic Phytophthora species, P. infestans, P. capsici, and P. nicotianae but not in A1 mating type strains. The response was concentration-dependent, and the oospores were viable. These results demonstrate that the biological activity of the synthetic hormone resembles that of the natural hormone α1. Mating hormones are essential components in the sexual life cycle of a variety of organisms. For plant pathogens like Phytophthora, sexual reproduction is important as a source of genetic variation. Moreover, the thick-walled oospores are the most durable propagules that can survive harsh environmental conditions. Sexual reproduction can thus greatly affect disease epidemics. The availability of synthetic compounds mimicking the activity of Phytophthora mating hormone will be instrumental for further unravelling sexual reproduction in this important group of plant pathogens.

Anthropogenic impact on amorphous silica pools in temperate soils

Human land use changes perturb biogeochemical silica (Si) cycling in terrestrial ecosystems. This directly affects Si mobilisation and Si storage and influences Si export from the continents, although the magnitude of the impact is unknown. A major reason for our lack of understanding is that very little information exists on how land use affects amorphous silica (ASi) storage in soils. We have quantified and compared total alkali-extracted (PSi<sub>a</sub>) and easily soluble (PSi<sub>e</sub>) Si pools at four sites along a gradient of anthropogenic disturbance in southern Sweden. Land use clearly affects ASi pools and their distribution. Total PSi</sub>a</sub> and PSi<sub>e</sub> for a continuous forested site at Siggaboda Nature Reserve (66 900 ± 22 800 kg SiO<sub>2</sub> ha<sup>−1</sup> and 952 ± 16 kg SiO<sub>2</sub> ha<sup>−1</sup>) are significantly higher than disturbed land use types from the Råshult Culture Reserve including arable land (28 800 ± 7200 kg SiO<sub>2</sub> ha<sup>−1</sup> and 239 ± 91 kg SiO<sub>2</sub> ha<sup>−1</sup>), pasture sites (27 300 ± 5980 kg SiO<sub>2</sub> ha<sup>−1</sup> and 370 ± 129 kg SiO<sub>2</sub> ha<sup>−1</sup>) and grazed forest (23 600 ± 6370 kg SiO<sub>2</sub> ha<sup>−1</sup> and 346 ± 123 kg SiO<sub>2</sub> ha<sup>−1</sup>). Vertical PSi<sub>a</sub> and PSi<sub>e</sub> profiles show significant (<i>p</i> < 0.05) variation among the sites. These differences in size and distribution are interpreted as the long-term effect of reduced ASi replenishment, as well as changes in ecosystem specific pedogenic processes and increased mobilisation of the PSi<sub>a</sub> in disturbed soils. We have also made a first, though rough, estimate of the magnitude of change in temperate continental ASi pools due to human disturbance. Assuming that our data are representative, we estimate that total ASi storage in soils has declined by ca. 10 % since the onset of agricultural development (3000 BCE). Recent agricultural expansion (after 1700 CE) may have resulted in an average additional export of 1.1 ± 0.8 Tmol Si yr<sup>−1</sup> from the soil reservoir to aquatic ecosystems. This is ca. 20 % to the global land-ocean Si flux carried by rivers. It is necessary to update this estimate in future studies, incorporating differences in pedology, geology and climatology over temperate regions, but data are currently not sufficient. Yet, our results emphasize the importance of human activities for Si cycling in soils and for the land-ocean Si flux

Characterization of a wheat HSP70 gene and its expression in response to stripe rust infection and abiotic stresses

Members of the family of 70-kD heat shock proteins (HSP70 s) play various stress-protective roles in plants. In this study, a wheat HSP70 gene was isolated from a suppression subtractive hybridization (SSH) cDNA library of wheat leaves infected by Puccinia striiformis f. sp. tritici. The gene, that was designated as TaHSC70, was predicted to encode a protein of 690 amino acids, with a molecular mass of 73.54 KDa and a pI of 5.01. Further analysis revealed the presence of a conserved signature that is characteristic for HSP70s and phylogenetic analysis demonstrated that TaHSC70 is a homolog of chloroplast HSP70s. TaHSC70 mRNA was present in leaves of both green and etiolated wheat seedlings and in stems and roots. The transcript level in roots was approximately threefold less than in leaves but light–dark treatment did not charge TaHSC70 expression. Following heat shock of wheat seedlings at 40°C, TaHSC70 expression increased in leaves of etiolated seedlings but remained stable at the same level in green seedlings. In addition, TaHSC70 was differentially expressed during an incompatible and compatible interaction with wheat-stripe rust, and there was a transient increase in expression upon treatment with methyl jasmonate (MeJA) treatment. Salicylic acid (SA), ethylene (ET) and abscisic acid (ABA) treatments had no influence on TaHSC70 expression. These results suggest that TaHSC70 plays a role in stress-related responses, and in defense responses elicited by infection with stripe rust fungus and does so via a JA-dependent signal transduction pathway