Suppression of soil-borne plant pathogens

Soil borne plant pathogens considerably reduce crop yields worldwide and are difficult to control due to their ”masked” occurrence in the heterogeneous soil environment. This hampers the efficacy of chemical - and microbiological control agents. Outbreaks of crop diseases are not only dependent on the presence of pathogen propagules in the soil, but are also influenced by soil-related properties like physico-chemical characteristics, microbial activity and community composition. Strong competition for limited available carbon substrates restricts or prevents germination and pre-infective growth of pathogens. This competition can occur directly by rapid exploitation of substrates, so called resource competition, or indirectly via inhibitory secondary metabolites, called interference competition The overall effect of all competition based mechanisms and the abiotic environment on disease development is known as “general disease suppression” and is the sum of all factors that reduce disease. The aim of this thesis was to study different aspects of general disease suppression, in order to get more insight into the interplay between microbial communities, pathogen dynamics, and substrate availability in different agricultural soils. The first objective was to study the role of microbial volatile organic compounds in natural disease suppression in agricultural soils. In chapter 2 a series of simultaneous experiments were performed on a agricultural soil that received different management practises. We showed a strong correlation between root infection and -biomass production in a bioassay and the suppressive effects of microbial volatiles on the in vitro growth of the pathogen Pythium intermedium. No or weak volatile suppression coincided with significant lower root biomass and a higher disease index, whereas a strong volatile suppression related to high biomass and a low disease index. Furthermore, the composition of the original soil bacterial community showed a drastic shift due to the legacy effects of management practices, coinciding with the loss of volatile suppression. By comparing the emission profiles and the bacterial community composition of the differently managed soils, candidate inhibitory compounds and volatile producing bacterial groups could be identified. Altogether these results indicate that volatile organic compounds can have an important role in general disease suppression. To follow up on volatile suppression chapter 3 investigates the influence of soil-related (abiotic and biotic) variables on volatile mediated in vitro growth inhibition of different plant pathogens via an extensive soil survey including 50 Dutch arable agricultural fields. The volatile mediated suppression of three phylogenetic different soil borne pathogens (Rhizoctonia solani, Fusarium oxysporum and Pythium intermedium) was linked to a wide range of soil-related variables with univariate and multivariate regression models. The overall suppression of different pathogens was linked to microbial activity and organic substrates. However, different pathogens showed different sensitivity to volatile suppression. Furthermore, the soil-related factors corresponding to volatile mediated suppression were pathogen specific. In total, the results described in this chapter show that part of volatile suppression for a particular pathogen is based on general microbial activity, but our data shows as well that the individual response is pathogen specific. Chapter 4 explores the reservoir of potential plant pathogens harboring agricultural soils before the start of the growth season, together with the environmental drivers of this pool of pathogens. By investigating the pathogenic seedbank in relation to its environment we assessed which soil-related variables could explain differences among site pathogen community composition. Pathogens differing in phylogeny or mode of infection were related to different soil variables. For example the among-site differences in the presence of oomycetes could not be related to their environmental context. On the other hand the variation in root and shoot fungal pathogen community composition was linked to soil physico-chemical properties and non-pathogen microbial community composition, with potentially a significant role of litter saprophytes therein. As the presence of pathogen propagules in soil is not necessarily related to disease incidence, chapter 5 investigates the dynamics of root pathogens in the presence of a root in a model rhizosphere. We developed a qPCR based assay to test the growth response of a pathogen (Pythium intermedium) to the presence of root exudates over time. This exposure to root exudates showed soil specific pathogen dynamics. This finding may indicate that in situ (microbial) processes can successfully prevent pathogen development in some of the soils but not in others. Thus this method has the potential to provide an alternative way to assess the susceptibility of a soil to certain soil-borne diseases. The results of this thesis gave new insights into different aspects of disease suppression in agricultural soils which could serve as a fundament to develop environmentally-friendly control methods based on natural occurring ecological processes. Ideas for the implementation of this study and future research are discussed in chapter 6. </p

Soil pathogen-aphid interactions under differences in soil organic matter and mineral fertilizer

There is increasing evidence showing that microbes can influence plant-insect interactions. In addition, various studies have shown that aboveground pathogens can alter the interactions between plants and insects. However, little is known about the role of soil-borne pathogens in plant-insect interactions. It is also not known how environmental conditions, that steer the performance of soil-borne pathogens, might influence these microbe-plant-insect interactions. Here, we studied effects of the soil-borne pathogen Rhizoctonia solani on aphids (Sitobion avenae) using wheat (Triticum aestivum) as a host. In a greenhouse experiment, we tested how different levels of soil organic matter (SOM) and fertilizer addition influence the interactions between plants and aphids. To examine the influence of the existing soil microbiome on the pathogen effects, we used both unsterilized field soil and sterilized field soil. In unsterilized soil with low SOM content, R. solani addition had a negative effect on aphid biomass, whereas it enhanced aphid biomass in soil with high SOM content. In sterilized soil, however, aphid biomass was enhanced by R. solani addition and by high SOM content. Plant biomass was enhanced by fertilizer addition, but only when SOM content was low, or in the absence of R. solani. We conclude that belowground pathogens influence aphid performance and that the effect of soil pathogens on aphids can be more positive in the absence of a soil microbiome. This implies that experiments studying the effect of pathogens under sterile conditions might not represent realistic interactions. Moreover, pathogen-plant-aphid interactions can be more positive for aphids under high SOM conditions. We recommend that soil conditions should be taken into account in the study of microbe-plant-insect interactions

CRISPR/Cas9-induced (CTG⋅CAG)n repeat instability in the myotonic dystrophy type 1 locus: implications for therapeutic genome editing

Myotonic dystrophy type 1 (DM1) is caused by (CTG⋅CAG)n-repeat expansion within the DMPK gene and thought to be mediated by a toxic RNA gain of function. Current attempts to develop therapy for this disease mainly aim at destroying or blocking abnormal properties of mutant DMPK (CUG)n RNA. Here, we explored a DNA-directed strategy and demonstrate that single clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-cleavage in either its 5′ or 3′ unique flank promotes uncontrollable deletion of large segments from the expanded trinucleotide repeat, rather than formation of short indels usually seen after double-strand break repair. Complete and precise excision of the repeat tract from normal and large expanded DMPK alleles in myoblasts from unaffected individuals, DM1 patients, and a DM1 mouse model could be achieved at high frequency by dual CRISPR/Cas9-cleavage at either side of the (CTG⋅CAG)n sequence. Importantly, removal of the repeat appeared to have no detrimental effects on the expression of genes in the DM1 locus. Moreover, myogenic capacity, nucleocytoplasmic distribution, and abnormal RNP-binding behavior of transcripts from the edited DMPK gene were normalized. Dual sgRNA-guided excision of the (CTG⋅CAG)n tract by CRISPR/Cas9 technology is applicable for developing isogenic cell lines for research and may provide new therapeutic opportunities for patients with DM1

Cost of fouling in full-scale reverse osmosis and nanofiltration installations in the Netherlands

The economic impact of fouling in spiral wound membranes is not yet well explored. There has been an established assumption that the cost of fouling in membrane processes is significant, but this hypothesis has not been thoroughly evaluated. We conducted an economic analysis on seven full-scale installations, four nano filtration (NF) and three reverse osmosis (RO), to estimate the cost of fouling in industrial plants. The cost of fouling was calculated in detail, including costs of increase in feed channel pressure drop, water permeability reduction, early membrane replacement, and extensive cleaning-in-place (CIP). The estimated cost of fouling was expressed as a fraction of operational expenses (OPEX) for each plant and the major cost factors in fouling and CIP costs were identified. The selected NF plants were fed with anoxic ground water, while the feed water to RO plants was either surface water or municipal wastewater effluent. All the NF plants produce drinking water, while the RO plants produce demineralized water for industrial applications. We found that the cost of fouling in the RO plants was around 24% of OPEX, while the fouling related costs in NF cases was only around 11% due to the low biofouling potential of the anoxic ground water. The major factor in the cost of fouling is the early membrane replacement cost, followed by additional energy and with only a minor contribution from the cleaning costs. The down-time cost (caused by the interruption of water production during a CIP event) can be the major CIP cost factor for the plants with frequent cleaning events, while the cost of chemicals dominates in the plants with non-frequent CIP. In case of manual cleaning-in-place, the cost of fouling is increased by around 2% for the RO plants with frequent CIP. The manual execution of CIP cleaning is an attention point to reconsider, as the reviewed plants hold an automated CIP cleaning, providing membrane productivity advantages

Bacteriën en schimmels op melkveebedrijf in beeld

Environmental Biolog

Microbiota in dung and milk differ between organic and conventional dairy farms

Organic farming is increasingly promoted as a means to reduce the environmental impact of artificial fertilizers, pesticides, herbicides, and antibiotics in conventional dairy systems. These factors potentially affect the microbial communities of the production stages (soil, silage, dung, and milk) of the entire farm cycle. However, understanding whether the microbiota representative of different production stages reflects different agricultural practices - such as conventional versus organic farming - is unknown. Furthermore, the translocation of the microbial community across production stages is scarcely studied. We sequenced the microbial communities of soil, silage, dung, and milk samples from organic and conventional dairy farms in the Netherlands. We found that community structure of soil fungi and bacteria significantly differed among soil types, but not between organic versus conventional farming systems. The microbial communities of silage also did not differ among conventional and organic systems. Nevertheless, the dung microbiota of cows and the fungal communities in the milk were significantly structured by agricultural practice. We conclude that, while the production stages of dairy farms seem to be disconnected in terms of microbial transfer, certain practices specific for each agricultural system, such as the content of diet and the use of antibiotics, are potential drivers of shifts in the cow's microbiota, including the milk produced. This may reflect differences in farm animal health and quality of dairy products depending on farming practices.Environmental Biolog

Prediabetes Is Associated With Structural Brain Abnormalities:The Maastricht Study

OBJECTIVE Structural brain abnormalities are key risk factors for brain diseases, such as dementia, stroke, and depression, in type 2 diabetes. It is unknown whether structural brain abnormalities already occur in prediabetes. Therefore, we investigated whether both prediabetes and type 2 diabetes are associated with lacunar infarcts (LIs), white matter hyperintensities (WMHs), cerebral microbleeds (CMBs), and brain atrophy. RESEARCH DESIGN and METHODS We used data from 2,228 participants (1,373 with normal glucose metabolism [NGM], 347 with prediabetes, and 508 with type 2 diabetes (oversampled); mean age 59.2 6 8.2 years; 48.3% women) of the Maastricht Study, a population-based cohort study. Diabetes status was determined with an oral glucose tolerance test. Brain imaging was performed with 3 Tesla MRI. Results were analyzed with multivariable logistic and linear regression analyses. RESULTS Prediabetes and type 2 diabetes were associated with the presence of LIs (odds ratio 1.61 [95% CI 0.98-2.63] and 1.67 [1.04-2.68], respectively; P trend = 0.027), larger WMH (b 0.07 log10-transformed mL [log-mL] [95% CI 0.00-0.15] and 0.21 log-mL [0.14-0.28], respectively; P trend <0.001), and smaller white matter volumes (b 24.0 mL [27.3 to 20.6] and 27.2 mL [210.4 to 24.0], respectively; P trend <0.001) compared with NGM. Prediabetes was not associated with gray matter volumes or the presence of CMBs. CONCLUSIONS Prediabetes is associated with structural brain abnormalities, with further deterioration in type 2 diabetes. These results indicate that, in middle-aged populations, structural brain abnormalities already occur in prediabetes, which may suggest that the treatment of early dysglycemia may contribute to the prevention of brain diseases

Recovery in the myogenic program of congenital myotonic dystrophy myoblasts after excision of the expanded (CTG)n repeat

The congenital form of myotonic dystrophy type 1 (cDM) is caused by the large-scale expansion of a (CTG•CAG)n repeat in DMPK and DM1-AS. The production of toxic transcripts with long trinucleotide tracts from these genes results in impairment of the myogenic differentiation capacity as cDM’s most prominent morpho-phenotypic hallmark. In the current in vitro study, we compared the early differentiation programs of isogenic cDM myoblasts with and without a (CTG)2600 repeat obtained by gene editing. We found that excision of the repeat restored the ability of cDM myoblasts to engage in myogenic fusion, preventing the ensuing myotubes from remaining immature. Although the cDM-typical epigenetic status of the DM1 locus and the expression of genes therein were not altered upon removal of the repeat, analyses at the transcriptome and proteome level revealed that early abnormalities in the temporal expression of differentiation regulators, myogenic progression markers, and alternative splicing patterns before and immediately after the onset of differentiation became normalized. Our observation that molecular and cellular features of cDM are reversible in vitro and can be corrected by repeat-directed genome editing in muscle progenitors, when already committed and poised for myogenic differentiation, is important information for the future development of gene therapy for different forms of myotonic dystrophy type 1 (DM1)

STXBP1 promotes Weibel-Palade body exocytosis through its interaction with the Rab27A effector Slp4-a.

Vascular endothelial cells contain unique rod-shaped secretory organelles, called Weibel-Palade bodies (WPBs), which contain the hemostatic protein von Willebrand factor (VWF) and a cocktail of angiogenic and inflammatory mediators. We have shown that the Rab27A effector synaptotagmin-like protein 4-a (Slp4-a) plays a critical role in regulating hormone-evoked WPB exocytosis. Using a nonbiased proteomic screen for targets for Slp4-a, we now identify syntaxin-binding protein 1 (STXBP1) and syntaxin-2 and -3 as endogenous Slp4-a binding partners in endothelial cells. Coimmunoprecipitations showed that STXBP1 interacts with syntaxin-2 and -3, but not with syntaxin-4. Small interfering RNA-mediated silencing of STXBP1 expression impaired histamine- and forskolin-induced VWF secretion. To further substantiate the role of STXBP1, we isolated blood outgrowth endothelial cells (BOECs) from an early infantile epileptic encephalopathy type 4 (EIEE4) patient carrying a de novo mutation in STXBP1. STXBP1-haploinsufficient EIEE4 BOECs contained similar numbers of morphologically normal WPBs compared with control BOECs of healthy donors; however, EIEE4 BOECs displayed significantly impaired histamine- and forskolin-stimulated VWF secretion. Based on these findings, we propose that the Rab27A-Slp4-a complex on WPB promotes exocytosis through an interaction with STXBP1, thereby controlling the release of vaso-active substances in the vasculature